Tyrosine kinase inhibitors containing a zinc binding moiety

ABSTRACT

The present invention relates to tyrosine kinase inhibitors that contain a zinc-binding moiety and their use in the treatment of tyrosine related diseases and disorders such as cancer. The said derivatives may further act as HDAC inhibitors.

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.13/010,542, filed Jan. 20, 2011, which is a divisional of U.S.application Ser. No. 11/852,450, filed Sep. 10, 2007 (now U.S. Pat. No.7,888,361), which claims the benefit of U.S. Provisional Application No.60/843,730, filed on Sep. 11, 2006 and U.S. Provisional Application No.60/895,901, filed on Mar. 20, 2007. The entire teachings of the aboveapplications are incorporated herein by reference.

BACKGROUND OF THE INVENTION

Protein kinases (PK) are enzymes that catalyze the phosphorylation ofhydroxyl groups of tyrosine, serine, and threonine residues of proteins.Many aspects of cell life such as cell growth, differentiation,proliferation, cell cycle and survival, depend on protein kinaseactivities. Furthermore, abnormal protein kinase activity has beenrelated to a host of disorders such as cancer and inflammation.Therefore, there is a great deal of effort directed to identifying waysto modulate protein kinase activities.

Growth factor receptors with PTK activity are known as receptor tyrosinekinases (“RTKs”). They comprise a large family of transmembranereceptors with diverse biological activity. At present, at leastnineteen distinct subfamilies of RTKs have been identified. Examples ofRTKs subfamily include VEGFR-1, VEGFR-2, Flt-3 c-Kit and PDGFR.

In addition to the RTKs, there also exists a family of entirelyintracellular PTKs called “non-receptor tyrosine kinases” or “cellulartyrosine kinases”. This latter designation, abbreviated “CTK”, will beused herein. CTKs do not contain extracellular and transmembranedomains. At present, over 24 CTKs in 11 subfamilies (Src, Frk, Btk, Csk,Abl, Zap70, Fes/Fps, Fak, Jak and Ack) have been identified. The Srcsubfamily appear so far to be the largest group of CTKs and includesSrc, Yes, Fyn, Lyn, Lck, Blk, Hck, Fgr and Yrk. For a more detaileddiscussion of CTKs, see Bolen, Oncogene, 1993, 8:2025-2031.

Chronic myeloid leukaemia (CML) is a stem cell disease characterized byan increased production and accumulation of clonal BCR-ABL-positivecells in haematopoietic tissues. Treatment of CML has been greatlyenhanced by the development of Imatinib mesylate, a specific inhibitorof the BCR-ABL tyrosine kinase. However, relapse occurs, mainly as aresult of the outgrowth of leukemic subclones with imatinib-resistantBCR-ABL mutations. Dasatinib, an second generation BCR-ABL inhibitorthat targets most imatinib-resistant BCR-ABL mutations, induceshematologic and cytogenetic responses in patients with CML orPh-positive ALL who cannot tolerate or are resistant to imatinib. Inaddition to inhibiting BCR-ABL, dasatinib has been reported to blockactivities of the SFKs, Lyn and Src on human prostate cancer cells (Nam,S., et al., Cancer Res 2005, 2005, 65 (20), 9185-9188.

Elucidation of the complex and multifactorial nature of various diseasesthat involve multiple pathogenic pathways and numerous molecularcomponents suggests that multi-targeted therapies may be advantageousover mono-therapies. Recent combination therapies with two or moreagents for many such diseases in the areas of oncology, infectiousdisease, cardiovascular disease and other complex pathologiesdemonstrate that this combinatorial approach may provide advantages withrespect to overcoming drug resistance, reduced toxicity and, in somecircumstances, a synergistic therapeutic effect compared to theindividual components.

Certain cancers have been effectively treated with such a combinatorialapproach; however, treatment regimes using a cocktail of cytotoxic drugsoften are limited by dose limiting toxicities and drug-druginteractions. More recent advances with molecularly targeted drugs haveprovided new approaches to combination treatment for cancer, allowingmultiple targeted agents to be used simultaneously, or combining thesenew therapies with standard chemotherapeutics or radiation to improveoutcome without reaching dose limiting toxicities. However, the abilityto use such combinations currently is limited to drugs that showcompatible pharmacologic and pharmacodynamic properties. In addition,the regulatory requirements to demonstrate safety and efficacy ofcombination therapies can be more costly and lengthy than correspondingsingle agent trials. Once approved, combination strategies may also beassociated with increased costs to patients, as well as decreasedpatient compliance owing to the more intricate dosing paradigmsrequired.

In the field of protein and polypeptide-based therapeutics it has becomecommonplace to prepare conjugates or fusion proteins that contain mostor all of the amino acid sequences of two differentproteins/polypeptides and that retain the individual binding activitiesof the separate proteins/polypeptides. This approach is made possible byindependent folding of the component protein domains and the large sizeof the conjugates that permits the components to bind their cellulartargets in an essentially independent manner. Such an approach is not,however, generally feasible in the case of small molecule therapeutics,where even minor structural modifications can lead to major changes intarget binding and/or the pharmacokinetic/pharmacodynamic properties ofthe resulting molecule.

Histone acetylation is a reversible modification, with deacetylationbeing catalyzed by a family of enzymes termed histone deacetylases(HDACs). HDAC's are represented by X genes in humans and are dividedinto four distinct classes (J Mol Biol, 2004, 338:1, 17-31). Inmammalians class I HDAC's (HDAC1-3, and HDAC8) are related to yeast RPD3HDAC, class 2 (HDAC4-7, HDAC9 and HDAC10) related to yeast HDA1, class 4(HDAC11), and class 3 (a distinct class encompassing the sirtuins whichare related to yeast Sir2).

Csordas, Biochem. J., 1990, 286: 23-38 teaches that histones are subjectto post-translational acetylation of the, ε-amino groups of N-terminallysine residues, a reaction that is catalyzed by histone acetyltransferase (HAT1). Acetylation neutralizes the positive charge of thelysine side chain, and is thought to impact chromatin structure. Indeed,access of transcription factors to chromatin templates is enhanced byhistone hyperacetylation, and enrichment in underacetylated histone H4has been found in transcriptionally silent regions of the genome(Taunton et al., Science, 1996, 272:408-411). In the case of tumorsuppressor genes, transcriptional silencing due to histone modificationcan lead to oncogenic transformation and cancer.

Several classes of HDAC inhibitors currently are being evaluated byclinical investigators. The first FDA approved HDAC inhibitor isSuberoylanilide hydroxamic acid (SAHA, ZOLINZA®) for the treatment ofcutaneous T-cell lymphoma (CTCL). Other HDAC inhibitors includehydroxamic acid derivatives, PXD101 and LAQ824, are currently in theclinical development. In the benzamide class of HDAC inhibitors, MS-275,MGCD0103 and CI-994 have reached clinical trials. Mourne et al.(Abstract #4725, AACR 2005), demonstrate that thiophenyl modification ofbenzamides significantly enhance HDAC inhibitory activity against HDAC1.

Recent advances suggest that HDAC inhibitors in combination with othertargeted agents may provide advantageous results in the treatment ofcancer. For example, co-treatment with SAHA significantly increasedEGFR2 antibody trastuzumab-induced apoptosis of BT-474 and SKBR-3 cellsand induced synergistic cytotoxic effects against the breast cancercells (Bali, Clin. Cancer Res., 2005, 11, 3392). HDAC inhibitors, suchas SAHA, have demonstrated synergistic antiproliferative and apoptoticeffects when used in combination with gefitinib in head and neck cancercell lines, including lines that are resistant to gefitinib monotherapy(Bruzzese et al., Proc. AACR, 2004). Pretreating gefitinib resistantcell lines with the HDAC inhibitor, MS-275, led to a growth-inhibitoryand apoptotic effect of gefitinib similar to that seen ingefitinib-sensitive NSCLC cell lines including those harboring EGFRmutations (Witta S. E., et al., Cancer Res, 2006, 66:2, 944-50). TheHDAC inhibitor PXD101 has been shown to act synergistically to inhibitproliferation with the EGFR1 inhibitor TARCEVA® (erlotinib)(WO2006082428A2). Synergy between NVP-LAQ824 and imatinib mesylate wasdemonstrated against BCR/ABL-expressing K562 myeloid leukemia celllines. (Weisberg et al., Leukemia. 2004, 18, 1951).

Current therapeutic regimens of the types described above attempt toaddress the problem of drug resistance by the administration of multipleagents. However, the combined toxicity of multiple agents due tooff-target side effects as well as drug-drug interactions often limitthe effectiveness of this approach. Moreover, it often is difficult tocombine compounds having differing pharmacokinetics into a single dosageform, and the consequent requirement of taking multiple medications atdifferent time intervals leads to problems with patient compliance thatcan undermine the efficacy of the drug combinations. In addition, thehealth care costs of combination therapies may be greater than forsingle molecule therapies. Moreover, it may be more difficult to obtainregulatory approval of a combination therapy since the burden fordemonstrating activity/safety of a combination of two agents may begreater than for a single agent. (Dancey J & Chen H, Nat. Rev. DrugDis., 2006, 5:649). The development of novel agents that target multipletherapeutic targets selected not by virtue of cross reactivity, butthrough rational design will help improve patient outcome while avoidingthese limitations. Thus, enormous efforts are still directed to thedevelopment of selective anti-cancer drugs as well as to new and moreefficacious combinations of known anti-cancer drugs.

SUMMARY OF THE INVENTION

The present invention relates to BCR-ABL kinase inhibitors that containa zinc-binding moiety and their use in the treatment of BCR-ABL relateddiseases and disorders such as cancer.

The compounds of the present invention may further act as HDAC or matrixmetalloproteinase (MMP) inhibitors by virtue of their ability to bindzinc ions. Surprisingly these compounds are active at multipletherapeutic targets and are effective for treating disease. Moreover, insome cases it has even more surprisingly been found that the compoundshave enhanced activity when compared to the activities of combinationsof separate molecules individually having the BCR-ABL and HDACactivities. In other words, the combination of pharmacophores into asingle molecule may provide a synergistic effect as compared to theindividual pharmacophores. More specifically, it has been found that itis possible to prepare compounds that simultaneously contain a firstportion of the molecule that binds zinc ions and thus permits inhibitionof HDAC and/or matrix metalloproteinase (MMP) activity and at least asecond portion of the molecule that permits binding to a separate anddistinct target that inhibits BCR-ABL and thus provides therapeuticbenefit. Preferably, the compounds of the present invention inhibit bothBCR-ABL and HDAC activity.

Accordingly, the present invention provides a compound having thegeneral formulae (I) and (II):

or its geometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof,wherein

-   -   Cz is selected from aryl, substituted aryl, heteroaryl,        substituted heteroaryl, heterocylic and substituted        heterocyclic;    -   Ar is aryl, substituted aryl heteroaryl or substituted        heteroaryl;    -   X₃ is NH, alkylamino, O or S;    -   Z₂ is O, S, NH or alkylamino;    -   Y₂ is N or CR₂₀; where R₂₀ is selected from hydrogen, halogen,        aliphatic, substituted aliphatic, aryl, substituted aryl,        heteroaryl, substituted heteroaryl;    -   R₂₁ is hydrogen or aliphatic;    -   B is a direct bond or straight- or branched-, substituted or        unsubstituted alkyl, substituted or unsubstituted alkenyl,        substituted or unsubstituted alkynyl, arylalkyl, arylalkenyl,        arylalkynyl, heteroarylalkyl, heteroarylalkenyl,        heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl,        heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl,        cycloalkenyl, alkylarylalkyl, alkylarylalkenyl,        alkylarylalkynyl, alkenylarylalkyl, alkenylarylalkenyl,        alkenylarylalkynyl, alkynylarylalkyl, alkynylarylalkenyl,        alkynylarylalkynyl, alkylheteroarylalkyl,        alkylheteroarylalkenyl, alkylheteroarylalkynyl,        alkenylheteroarylalkyl, alkenylheteroarylalkenyl,        alkenylheteroarylalkynyl, alkynylheteroarylalkyl,        alkynylheteroarylalkenyl, alkynylheteroarylalkynyl,        alkylheterocyclylalkyl, alkylheterocyclylalkenyl,        alkylhererocyclylalkynyl, alkenylheterocyclylalkyl,        alkenylheterocyclylalkenyl, alkenylheterocyclylalkynyl,        alkynylheterocyclylalkyl, alkynylheterocyclylalkenyl,        alkynylheterocyclylalkynyl, alkylaryl, alkenylaryl, alkynylaryl,        alkylheteroaryl, alkenylheteroaryl, or alkynylhereroaryl, which        one or more methylenes can be interrupted or terminated by O, S,        S(O), SO₂, N(R₈), C(O), substituted or unsubstituted aryl,        substituted or unsubstituted heteroaryl, substituted or        unsubstituted heterocyclic; where R₈ is hydrogen, acyl,        aliphatic or substituted aliphatic.

In one embodiment, the linker B is between 1-24 atoms, preferably 4-24atoms, preferably 4-18 atoms, more preferably 4-12 atoms, and mostpreferably about 4-10 atoms.

C is selected from:

where W is O or S; Y is absent, N, or CH; Z is N or CH; R₇ and R₉ areindependently hydrogen, OR′, aliphatic or substituted aliphatic, whereinR′ is hydrogen, aliphatic, substituted aliphatic or acyl; provided thatif R₇ and R₉ are both present, one of R₇ or R₉ must be OR′ and if Y isabsent, R₉ must be OR′; and R₂₁ is hydrogen, acyl, aliphatic, orsubstituted aliphatic;

where W is O or S; J is O, NH or NCH₃; and R₁₀ is hydrogen or loweralkyl;

where W is O or S; Y₁ and Z₁ are independently N, C or CH; and

where Z, Y, and W are as previously defined; R₁₁ and R₁₂ areindependently selected from hydrogen or aliphatic; R₁, R₂ and R₃ areindependently selected from hydrogen, hydroxy, amino, halogen, alkoxy,substituted alkoxy, alkylamino, substituted alkylamino, dialkylamino,substituted dialkylamino, substituted or unsubstituted alkylthio,substituted or unsubstituted alkylsulfonyl, CF₃, CN, N₃, NO₂, sulfonyl,acyl, aliphatic, substituted aliphatic, aryl, substituted aryl,heteroaryl, substituted heteroaryl, heterocyclic, and substitutedheterocyclic.

DETAILED DESCRIPTION OF THE INVENTION

In a first embodiment of the compounds of the present invention arecompounds represented by formulae (I) and (II) as illustrated above, orits geometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof.

In a second embodiment of the compounds of the present invention arecompounds represented by formula (III) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein R₄ is hydrogen, aliphatic or substituted aliphatic; C, B, Y₂,Z₂, Ar and R₂₁ are as previously defined.

In a third embodiment of the compounds of the present invention arecompounds represented by formula (IV) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein n is 1-9; R′, Z₂, Ar and R₂₁ are as previously defined.

In a fourth embodiment of the compounds of the present invention arecompounds represented by formula (V) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein m and n are independently 1-9; R₁₈ is independently R₈; R′, Z₂,Ar R₈, and R₂₁ are as previously defined.

In a fifth embodiment of the compounds of the present invention arecompounds represented by formula (VI) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein m is 0-9; W is O, NH, alkylamino, S, SO, SO₂; Z₂, Ar R′ R₈, andR₂₁ are as previously defined.

In a sixth embodiment of the compounds of the present invention arecompounds represented by formula (VII) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein M₁ is absent, O, NH, alkylamino, or C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl; M₂ is absent, C₁-C₆ alkyl, O, NH, alkylamine, S, SO, SO₂,heterocyclic, heteroaryl, aryl, or C≡O; M₃ is absent, O, NH, alkyamino,C═O, C₁-C₆ alkyl, C₂-C₆ alkenyl, or C₂-C₆ alkynyl; M₄ is absent, O, NH,alkyamino, heterocyclic, heteroaryl or aryl; M₅ is absent, C₁-C₈ alkyl,C₂-C₈ alkenyl, C₂-C₈ alkynyl, heterocyclic, heteroaryl or aryl; R′, R₁,Z₂, Ar and R₂₁ are as previously defined.

In a seventh embodiment of the compounds of the present invention arecompounds represented by formula (VIII) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein M₁ is absent, O, NH, alkylamino, or C₁-C₆ alkyl, C₂-C₆ alkenyl,C₂-C₆ alkynyl; M₂ is absent, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl,O, NH, alkylamine, S, SO, SO₂, heterocyclic, heteroaryl, aryl or C═O; M₃is absent, O, NH, alkyamino, S, SO, SO₂, C₁-C₈ alkyl, C₂-C₈ alkenyl,C₂-C₈ alkynyl, heterocyclic, heteroaryl, aryl or C═O; M₄ is absent, O,NH, alkyamino, CO, S, SO, SO₂, heterocyclic, heteroaryl or aryl; M₅ isabsent, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, heterocyclic,heteroaryl or aryl; R′, Z₂, Ar and R₂₁ are as previously defined.

In an eighth embodiment of the compounds of the present invention arecompounds represented by formula (IX) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

-   -   Cz is selected from aryl, substituted aryl, heteroaryl,        substituted heteroaryl, heterocylic and substituted        heterocyclic;    -   Ar is aryl, substituted aryl heteroaryl or substituted        heteroaryl;    -   X₃ is NH, O or S;    -   Z₂ is O, S, or NH;    -   Y₂ is N or CR₂₀; where R₂₀ is selected from hydrogen, halogen,        aliphatic, substituted aliphatic, aryl, substituted aryl,        heteroaryl, substituted heteroaryl;    -   R₈ is hydrogen or aliphatic;    -   B is a direct bond or straight- or branched-, substituted or        unsubstituted alkyl, substituted or unsubstituted alkenyl,        substituted or unsubstituted alkynyl, arylalkyl, arylalkenyl,        arylalkynyl, heteroarylalkyl, heteroarylalkenyl,        heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl,        heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl,        alkylarylalkyl, alkylarylalkenyl, alkylarylalkynyl,        alkenylarylalkyl, alkenylarylalkenyl, alkenylarylalkynyl,        alkynylarylalkyl, alkynylarylalkenyl, alkynylarylalkynyl,        alkylheteroarylalkyl, alkylheteroarylalkenyl,        alkylheteroarylalkynyl, alkenylheteroarylalkyl,        alkenylheteroarylalkenyl, alkenylheteroarylalkynyl,        alkynylheteroarylalkyl, alkynylheteroarylalkenyl,        alkynylheteroarylalkynyl, alkylheterocyclylalkyl,        alkylheterocyclylalkenyl, alkylhererocyclylalkynyl,        alkenylheterocyclylalkyl, alkenylheterocyclylalkenyl,        alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl,        alkynylheterocyclylalkenyl, or alkynylheterocyclylalkynyl, which        one or more methylenes can be interrupted or terminated by O, S,        S(O), SO₂, N(R₈), C(O), substituted or unsubstituted aryl,        substituted or unsubstituted heteroaryl, substituted or        unsubstituted heterocyclic; where R₈ is hydrogen or aliphatic        group;    -   C is selected from:

where W is O or S; Y is absent, N, or CH; Z is N or CH; R₇ and R₉ areindependently hydrogen, hydroxy, aliphatic group, provided that if R₇and R₉ are both present, one of R₇ or R₉ must be hydroxy and if Y isabsent, R₉ must be hydroxy; and R₈ is hydrogen or aliphatic group;

where W is O or S; J is O, NH or NCH₃; and R₁₀ is hydrogen or loweralkyl;

where W is O or S; Y₁ and Z₁ are independently N, C or CH; and

where Z, Y, and W are as previously defined; R₁₁ and R₁₂ areindependently selected from hydrogen or aliphatic; R₁, R₂ and R₃ areindependently selected from hydrogen, hydroxy, amino, halogen, alkoxy,alkylamino, dialkylamino, CF₃, CN, NO₂, sulfonyl, acyl, aliphatic,substituted aliphatic, aryl, substituted aryl, heteroaryl, substitutedheteroaryl, heterocyclic, and substituted heterocyclic.

In a ninth embodiment of the compounds of the present invention arecompounds represented by formula (X) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein R₄ is hydrogen, aliphatic or substituted aliphatic; C, B, Y₂,Z₂, Ar and R₈ are as previously defined.

In a tenth embodiment of the compounds of the present invention arecompounds represented by formula (XI) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein n is 1-9; Z₂, Ar and R₈ are as previously defined.

In an eleventh embodiment of the compounds of the present invention arecompounds represented by formula (XII) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein n is 1-9; Z₂, Ar and R₈ are as previously defined.

In a twelfth embodiment of the compounds of the present invention arecompounds represented by formula (XIII) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein m is 0-9; Z₂, Ar and R₈ are as previously defined.

In a thirteenth embodiment of the compounds of the present invention arecompounds represented by formula (XIV) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein m is 0-9; Z₂, Ar and R₈ are as previously defined.

In a fourteenth embodiment of the compounds of the present invention arecompounds represented by formula (XV) as illustrated below, or itsgeometric isomers, enantiomers, diastereomers, racemates,pharmaceutically acceptable salts, prodrugs and solvates thereof:

wherein m is 0-9; Z₂, Ar and R₈ are as previously defined.

Representative compounds according to the invention are those selectedfrom the Table A below or its geometric isomers, enantiomers,diastereomers, racemates, pharmaceutically acceptable salts, prodrugsand solvates thereof:

TABLE A Compound # Structure 1

2

3

4

5

6

7

8

9

10

11

12

13

14

15

16

17

18

19

20

21

22

23

24

25

26

27

28

29

30

31

32

33

34

35

36

37

38

39

40

41

42

43

The invention further provides methods for the prevention or treatmentof diseases or conditions involving aberrant proliferation,differentiation or survival of cells. In one embodiment, the inventionfurther provides for the use of one or more compounds of the inventionin the manufacture of a medicament for halting or decreasing diseasesinvolving aberrant proliferation, differentiation, or survival of cells.In preferred embodiments, the disease is cancer. In one embodiment, theinvention relates to a method of treating cancer in a subject in need oftreatment comprising administering to said subject a therapeuticallyeffective amount of a compound of invention.

The term “cancer” refers to any cancer caused by the proliferation ofmalignant neoplastic cells, such as tumors, neoplasms, carcinomas,sarcomas, leukemias, lymphomas and the like. For example, cancersinclude, but are not limited to, mesothelioma, leukemias and lymphomassuch as cutaneous T-cell lymphomas (CTCL), noncutaneous peripheralT-cell lymphomas, lymphomas associated with human T-cell lymphotrophicvirus (HTLV) such as adult T-cell leukemia/lymphoma (ATLL), B-celllymphoma, acute nonlymphocytic leukemias, chronic lymphocytic leukemia,chronic myelogenous leukemia, acute myelogenous leukemia, lymphomas, andmultiple myeloma, non-Hodgkin lymphoma, acute lymphatic leukemia (ALL),chronic lymphatic leukemia (CLL), Hodgkins lymphoma, Burkitt lymphoma,adult T-cell leukemia lymphoma, acute-myeloid leukemia (AML), chronicmyeloid leukemia (CML), or hepatocellular carcinoma. Further examplesinclude myelodisplastic syndrome, childhood solid tumors such as braintumors, neuroblastoma, retinoblastoma, Wilms' tumor, bone tumors, andsoft-tissue sarcomas, common solid tumors of adults such as head andneck cancers (e.g., oral, laryngeal, nasopharyngeal and esophageal),genitourinary cancers (e.g., prostate, bladder, renal, uterine, ovarian,testicular), lung cancer (e.g., small-cell and non small cell), breastcancer, pancreatic cancer, melanoma and other skin cancers, stomachcancer, brain tumors, tumors related to Gorlin's syndrome (e.g.,medulloblastoma, meningioma, etc.), and liver cancer. Additionalexemplary forms of cancer which may be treated by the subject compoundsinclude, but are not limited to, cancer of skeletal or smooth muscle,stomach cancer, cancer of the small intestine, rectum carcinoma, cancerof the salivary gland, endometrial cancer, adrenal cancer, anal cancer,rectal cancer, parathyroid cancer, and pituitary cancer.

Additional cancers that the compounds described herein may be useful inpreventing, treating and studying are, for example, colon carcinoma,familiary adenomatous polyposis carcinoma and hereditary non-polyposiscolorectal cancer, or melanoma. Further, cancers include, but are notlimited to, labial carcinoma, larynx carcinoma, hypopharynx carcinoma,tongue carcinoma, salivary gland carcinoma, gastric carcinoma,adenocarcinoma, thyroid cancer (medullary and papillary thyroidcarcinoma, renal carcinoma, kidney parenchyma carcinoma, cervixcarcinoma, uterine corpus carcinoma, endometrium carcinoma, chorioncarcinoma, testis carcinoma, urinary carcinoma, melanoma, brain tumorssuch as glioblastoma, astrocytoma, meningioma, medulloblastoma andperipheral neuroectodermal tumors, gall bladder carcinoma, bronchialcarcinoma, multiple myeloma, basalioma, teratoma, retinoblastoma,choroidea melanoma, seminoma, rhabdomyosarcoma, craniopharyngeoma,osteosarcoma, chondrosarcoma, myosarcoma, liposarcoma, fibrosarcoma,Ewing sarcoma, and plasmocytoma. In another aspect of the invention, thepresent invention provides for the use of one or more compounds of theinvention in the manufacture of a medicament for the treatment ofcancer.

In one embodiment, the present invention includes the use of one or morecompounds of the invention in the manufacture of a medicament thatprevents further aberrant proliferation, differentiation, or survival ofcells. For example, compounds of the invention may be useful inpreventing tumors from increasing in size or from reaching a metastaticstate. The subject compounds may be administered to halt the progressionor advancement of cancer or to induce apoptosis or to inhibit tumorangiogenesis. In addition, the instant invention includes use of thesubject compounds to prevent a recurrence of cancer.

This invention further embraces the treatment or prevention of cellproliferative disorders such as hyperplasias, dysplasias andpre-cancerous lesions. Dysplasia is the earliest form of pre-cancerouslesion recognizable in a biopsy by a pathologist. The subject compoundsmay be administered for the purpose of preventing said hyperplasias,dysplasias or pre-cancerous lesions from continuing to expand or frombecoming cancerous. Examples of pre-cancerous lesions may occur in skin,esophageal tissue, breast and cervical intra-epithelial tissue.

“Combination therapy” includes the administration of the subjectcompounds in further combination with other biologically activeingredients (such as, but not limited to, a second and differentantineoplastic agent) and non-drug therapies (such as, but not limitedto, surgery or radiation treatment). For instance, the compounds of theinvention can be used in combination with other pharmaceutically activecompounds, preferably compounds that are able to enhance the effect ofthe compounds of the invention. The compounds of the invention can beadministered simultaneously (as a single preparation or separatepreparation) or sequentially to the other drug therapy. In general, acombination therapy envisions administration of two or more drugs duringa single cycle or course of therapy.

In one aspect of the invention, the subject compounds may beadministered in combination with one or more separate agents thatmodulate protein kinases involved in various disease states. Examples ofsuch kinases may include, but are not limited to: serine/threoninespecific kinases, receptor tyrosine specific kinases and non-receptortyrosine specific kinases. Serine/threonine kinases include mitogenactivated protein kinases (MAPK), meiosis specific kinase (MEK), RAF andaurora kinase. Examples of receptor kinase families include epidermalgrowth factor receptor (EGFR) (e.g. HER2/neu, HER3, HER4, ErbB, ErbB2,ErbB3, ErbB4, Xmrk, DER, Let23); fibroblast growth factor (FGF) receptor(e.g. FGF-R1, GFF-R2/BEK/CEK3, FGF-R3/CEK2, FGF-R4/TKF, KGF-R);hepatocyte growth/scatter factor receptor (HGFR) (e.g, MET, RON, SEA,SEX); insulin receptor (e.g. IGFI-R); Eph (e.g. CEK5, CEK8, EBK, ECK,EEK, EHK-1, EHK-2, ELK, EPH, ERK, HEK, MDK2, MDK5, SEK); Axl (e.g.Mer/Nyk, Rse); RET; and platelet-derived growth factor receptor (PDGFR)(e.g. PDGFα-R, PDGβ-R, CSF1-R/FMS, SCF-R/C-KIT, VEGF-R/FLT, NEK/FLK1,FLT3/FLK2/STK-1). Non-receptor tyrosine kinase families include, but arenot limited to, BCR-ABL (e.g. p43^(abl), ARG); BTK (e.g. ITK/EMT, TEC);CSK, FAK, FPS, JAK, SRC, BMX, FER, CDK and SYK.

In another aspect of the invention, the subject compounds may beadministered in combination with one or more separate agents thatmodulate non-kinase biological targets or processes. Such targetsinclude histone deacetylases (HDAC), DNA methyltransferase (DNMT), heatshock proteins (e.g. HSP90), and proteosomes.

In a preferred embodiment, subject compounds may be combined withantineoplastic agents (e.g. small molecules, monoclonal antibodies,antisense RNA, and fusion proteins) that inhibit one or more biologicaltargets such as Zolinza, Tarceva, Iressa, Tykerb, Gleevec, Sutent,Sprycel, Nexavar, Sorafinib, CNF2024, RG108, BMS387032, Affinitak,Avastin, Herceptin, Erbitux, AG24322, PD325901, ZD6474, PD184322,Obatodax, ABT737 and AEE788. Such combinations may enhance therapeuticefficacy over efficacy achieved by any of the agents alone and mayprevent or delay the appearance of resistant mutational variants.

In certain preferred embodiments, the compounds of the invention areadministered in combination with a chemotherapeutic agent.Chemotherapeutic agents encompass a wide range of therapeutic treatmentsin the field of oncology. These agents are administered at variousstages of the disease for the purposes of shrinking tumors, destroyingremaining cancer cells left over after surgery, inducing remission,maintaining remission and/or alleviating symptoms relating to the canceror its treatment. Examples of such agents include, but are not limitedto, alkylating agents such as mustard gas derivatives (Mechlorethamine,cylophosphamide, chlorambucil, melphalan, ifosfamide), ethylenimines(thiotepa, hexamethylmelanine), Alkylsulfonates (Busulfan), Hydrazinesand Triazines (Altretamine, Procarbazine, Dacarbazine and Temozolomide),Nitrosoureas (Carmustine, Lomustine and Streptozocin), Ifosfamide andmetal salts (Carboplatin, Cisplatin, and Oxaliplatin); plant alkaloidssuch as Podophyllotoxins (Etoposide and Tenisopide), Taxanes (Paclitaxeland Docetaxel), Vinca alkaloids (Vincristine, Vinblastine, Vindesine andVinorelbine), and Camptothecan analogs (Irinotecan and Topotecan);anti-tumor antibiotics such as Chromomycins (Dactinomycin andPlicamycin), Anthracyclines (Doxorubicin, Daunorubicin, Epirubicin,Mitoxantrone, Valrubicin and Idarubicin), and miscellaneous antibioticssuch as Mitomycin, Actinomycin and Bleomycin; anti-metabolites such asfolic acid antagonists (Methotrexate, Pemetrexed, Raltitrexed,Aminopterin), pyrimidine antagonists (5-Fluorouracil, Floxuridine,Cytarabine, Capecitabine, and Gemcitabine), purine antagonists(6-Mercaptopurine and 6-Thioguanine) and adenosine deaminase inhibitors(Cladribine, Fludarabine, Mercaptopurine, Clofarabine, Thioguanine,Nelarabine and Pentostatin); topoisomerase inhibitors such astopoisomerase I inhibitors (Ironotecan, topotecan) and topoisomerase IIinhibitors (Amsacrine, etoposide, etoposide phosphate, teniposide);monoclonal antibodies (Alemtuzumab, Gemtuzumab ozogamicin, Rituximab,Trastuzumab, Ibritumomab Tioxetan, Cetuximab, Panitumumab, Tositumomab,Bevacizumab); and miscellaneous anti-neoplastics such as ribonucleotidereductase inhibitors (Hydroxyurea); adrenocortical steroid inhibitor(Mitotane); enzymes (Asparaginase and Pegaspargase); anti-microtubuleagents (Estramustine); and retinoids (Bexarotene, Isotretinoin,Tretinoin (ATRA).

In certain preferred embodiments, the compounds of the invention areadministered in combination with a chemoprotective agent.Chemoprotective agents act to protect the body or minimize the sideeffects of chemotherapy. Examples of such agents include, but are notlimited to, amfostine, mesna, and dexrazoxane.

It will be appreciated that compounds of the invention can be used incombination with an immunotherapeutic agent. One form of immunotherapyis the generation of an active systemic tumor-specific immune responseof host origin by administering a vaccine composition at a site distantfrom the tumor. Various types of vaccines have been proposed, includingisolated tumor-antigen vaccines and anti-idiotype vaccines. Anotherapproach is to use tumor cells from the subject to be treated, or aderivative of such cells (reviewed by Schirrmacher et al. (1995) J.Cancer Res. Clin. Oncol. 121:487). In U.S. Pat. No. 5,484,596, Hanna Jr.et al. claim a method for treating a resectable carcinoma to preventrecurrence or metastases, comprising surgically removing the tumor,dispersing the cells with collagenase, irradiating the cells, andvaccinating the patient with at least three consecutive doses of about10⁷ cells.

It will be appreciated that the compounds of the invention mayadvantageously be used in conjunction with one or more other therapeuticagents. Examples of suitable agents for adjunctive therapy include a5HT₁ agonist, such as a triptan (e.g. sumatriptan or naratriptan); anadenosine A1 agonist; an EP ligand; an NMDA modulator, such as a glycineantagonist; a sodium channel blocker (e.g. lamotrigine); a substance Pantagonist (e.g. an NK₁ antagonist); a cannabinoid; acetaminophen orphenacetin; a 5-lipoxygenase inhibitor; a leukotriene receptorantagonist; a DMARD (e.g. methotrexate); gabapentin and relatedcompounds; a tricyclic antidepressant (e.g. amitryptilline); a neuronestabilising antiepileptic drug; a mono-aminergic uptake inhibitor (e.g.venlafaxine); a matrix metalloproteinase inhibitor; a nitric oxidesynthase (NOS) inhibitor, such as an iNOS or an nNOS inhibitor; aninhibitor of the release, or action, of tumour necrosis factor .alpha.;an antibody therapy, such as a monoclonal antibody therapy; an antiviralagent, such as a nucleoside inhibitor (e.g. lamivudine) or an immunesystem modulator (e.g. interferon); an opioid analgesic; a localanaesthetic; a stimulant, including caffeine; an H₂-antagonist (e.g.ranitidine); a proton pump inhibitor (e.g. omeprazole); an antacid (e.g.aluminium or magnesium hydroxide; an antiflatulent (e.g. simethicone); adecongestant (e.g. phenylephrine, phenylpropanolamine, pseudoephedrine,oxymetazoline, epinephrine, naphazoline, xylometazoline,propylhexedrine, or levo-desoxyephedrine); an antitussive (e.g. codeine,hydrocodone, carmiphen, carbetapentane, or dextramethorphan); adiuretic; or a sedating or non-sedating antihistamine.

In another aspect of the invention, the subject compounds areadministered in combination with radiation therapy. Radiation iscommonly delivered internally (implantation of radioactive material nearcancer site) or externally from a machine that employs photon (x-ray orgamma-ray) or particle radiation. Where the combination therapy furthercomprises radiation treatment, the radiation treatment may be conductedat any suitable time so long as a beneficial effect from the co-actionof the combination of the therapeutic agents and radiation treatment isachieved. For example, in appropriate cases, the beneficial effect isstill achieved when the radiation treatment is temporally removed fromthe administration of the therapeutic agents, perhaps by days or evenweeks.

Matrix metalloproteinases (MMPs) are a family of zinc-dependent neutralendopeptidases collectively capable of degrading essentially all matrixcomponents. Over 20 MMP modulating agents are in pharmaceutical develop,almost half of which are indicated for cancer. The University of Torontoresearchers have reported that HDACs regulate MMP expression andactivity in 3T3 cells. In particular, inhibition of HDAC by trichostatinA (TSA), which has been shown to prevent tumorigenesis and metastasis,decreases mRNA as well as zymographic activity of gelatinase A (MMP2;Type IV collagenase), a matrix metalloproteinase, which is itself,implicated in tumorigenesis and metastasis (Ailenberg M., Silverman M.,Biochem Biophys Res Commun. 2002, 298:110-115). Another recent articlethat discusses the relationship of HDAC and MMPs can be found in YoungD. A., et al., Arthritis Research & Therapy, 2005, 7: 503. Furthermore,the commonality between HDAC and MMPs inhibitors is their zinc-bindingfunctionality. Therefore, in another aspect of the invention, compoundsof the invention can be used as MMP inhibitors.

In one aspect, the compounds may also be used in the treatment of adisorder involving, relating to or, associated with dysregulation ofhistone deacetylase (HDAC).

There are a number of disorders that have been implicated by or known tobe mediated at least in part by HDAC activity, where HDAC activity isknown to play a role in triggering disease onset, or whose symptoms areknown or have been shown to be alleviated by HDAC inhibitors. Disordersof this type that would be expected to be amenable to treatment with thecompounds of the invention include the following but not limited to:Anti-proliferative disorders (e.g. cancers); Neurodegenerative diseasesincluding Huntington's Disease, Polyglutamine disease, Parkinson'sDisease, Alzheimer's Disease, Seizures, Striatonigral degeneration,Progressive supranuclear palsy, Torsion dystonia, Spasmodic torticollisand dyskinesis, Familial tremor, Gilles de la Tourette syndrome, DiffuseLewy body disease, Progressive supranuclear palsy, Pick's disease,intracerebral hemorrhage, Primary lateral sclerosis, Spinal muscularatrophy, Amyotrophic lateral sclerosis, Hypertrophic interstitialpolyneuropathy, Retinitis pigmentosa, Hereditary optic atrophy,Hereditary spastic paraplegia, Progressive ataxia and Shy-Dragersyndrome; Metabolic diseases including Type 2 diabetes; DegenerativeDiseases of the Eye including Glaucoma, Age-related maculardegeneration, Rubeotic glaucoma; Inflammatory diseases and/or Immunesystem disorders including Rheumatoid Arthritis (RA), Osteoarthritis,Juvenile chronic arthritis, Graft versus Host disease, Psoriasis,Asthma, Spondyloarthropathy, psoriasis, Crohn's Disease, inflammatorybowel disease Colitis Ulcerosa, Alcoholic hepatitis, Diabetes,Sjoegrens's syndrome, Multiple Sclerosis, Ankylosing spondylitis,Membranous glomerulopathy, Discogenic pain, Systemic LupusErythematosus; Disease involving angiogenesis including cancer,psoriasis, rheumatoid arthritis; Psychological disorders includingbipolar disease, schizophrenia, mania, depression and dementia;Cardiovascular Diseases including heart failure, restenosis andarteriosclerosis; Fibrotic diseases including liver fibrosis, cysticfibrosis and angiofibroma; Infectious diseases including Fungalinfections, such as Candida Albicans, Bacterial infections, Viralinfections, such as Herpes Simplex, Protozoal infections, such asMalaria, Leishmania infection, Trypanosoma brucei infection,Toxoplasmosis and coccidiosis and Haematopoietic disorders includingthalassemia, anemia and sickle cell anemia.

In one embodiment compounds of the invention can be used to induce orinhibit apoptosis, a physiological cell death process critical fornormal development and homeostasis. Alterations of apoptotic pathwayscontribute to the pathogenesis of a variety of human diseases. Compoundsof the invention, as modulators of apoptosis, will be useful in thetreatment of a variety of human diseases with aberrations in apoptosisincluding cancer (particularly, but not limited to, follicularlymphomas, carcinomas with p53 mutations, hormone dependent tumors ofthe breast, prostate and ovary, and precancerous lesions such asfamilial adenomatous polyposis), viral infections (including, but notlimited to, herpesvirus, poxvirus, Epstein-Barr virus, Sindbis virus andadenovirus), autoimmune diseases (including, but not limited to,systemic lupus, erythematosus, immune mediated glomerulonephritis,rheumatoid arthritis, psoriasis, inflammatory bowel diseases, andautoimmune diabetes mellitus), neurodegenerative disorders (including,but not limited to, Alzheimer's disease, AIDS-related dementia,Parkinson's disease, amyotrophic lateral sclerosis, retinitispigmentosa, spinal muscular atrophy and cerebellar degeneration), AIDS,myelodysplastic syndromes, aplastic anemia, ischemic injury associatedmyocardial infarctions, stroke and reperfusion injury, arrhythmia,atherosclerosis, toxin-induced or alcohol induced liver diseases,hematological diseases (including, but not limited to, chronic anemiaand aplastic anemia), degenerative diseases of the musculoskeletalsystem (including, but not limited to, osteoporosis and arthritis),aspirin-sensitive rhinosinusitis, cystic fibrosis, multiple sclerosis,kidney diseases, and cancer pain.

In another aspect, the invention provides the use of compounds of theinvention for the treatment and/or prevention of immune response orimmune-mediated responses and diseases, such as the prevention ortreatment of rejection following transplantation of synthetic or organicgrafting materials, cells, organs or tissue to replace all or part ofthe function of tissues, such as heart, kidney, liver, bone marrow,skin, cornea, vessels, lung, pancreas, intestine, limb, muscle, nervetissue, duodenum, small-bowel, pancreatic-islet-cell, includingxeno-transplants, etc.; to treat or prevent graft-versus-host disease,autoimmune diseases, such as rheumatoid arthritis, systemic lupuserythematosus, thyroiditis, Hashimoto's thyroiditis, multiple sclerosis,myasthenia gravis, type I diabetes uveitis, juvenile-onset orrecent-onset diabetes mellitus, uveitis, Graves disease, psoriasis,atopic dermatitis, Crohn's disease, ulcerative colitis, vasculitis,auto-antibody mediated diseases, aplastic anemia, Evan's syndrome,autoimmune hemolytic anemia, and the like; and further to treatinfectious diseases causing aberrant immune response and/or activation,such as traumatic or pathogen induced immune disregulation, includingfor example, that which are caused by hepatitis B and C infections, HIV,staphylococcus aureus infection, viral encephalitis, sepsis, parasiticdiseases wherein damage is induced by an inflammatory response (e.g.,leprosy); and to prevent or treat circulatory diseases, such asarteriosclerosis, atherosclerosis, vasculitis, polyarteritis nodosa andmyocarditis. In addition, the present invention may be used toprevent/suppress an immune response associated with a gene therapytreatment, such as the introduction of foreign genes into autologouscells and expression of the encoded product. Thus in one embodiment, theinvention relates to a method of treating an immune response disease ordisorder or an immune-mediated response or disorder in a subject in needof treatment comprising administering to said subject a therapeuticallyeffective amount of a compound of the invention.

In another aspect, the invention provides the use of compounds of theinvention in the treatment of a variety of neurodegenerative diseases, anon-exhaustive list of which is: I. Disorders characterized byprogressive dementia in the absence of other prominent neurologic signs,such as Alzheimer's disease; Senile dementia of the Alzheimer type; andPick's disease (lobar atrophy); II. Syndromes combining progressivedementia with other prominent neurologic abnormalities such as A)syndromes appearing mainly in adults (e.g., Huntington's disease,Multiple system atrophy combining dementia with ataxia and/ormanifestations of Parkinson's disease, Progressive supranuclear palsy(Steel-Richardson-Olszewski), diffuse Lewy body disease, andcorticodentatonigral degeneration); and B) syndromes appearing mainly inchildren or young adults (e.g., Hallervorden-Spatz disease andprogressive familial myoclonic epilepsy); III. Syndromes of graduallydeveloping abnormalities of posture and movement such as paralysisagitans (Parkinson's disease), striatonigral degeneration, progressivesupranuclear palsy, torsion dystonia (torsion spasm; dystonia musculorumdeformans), spasmodic torticollis and other dyskinesis, familial tremor,and Gilles de la Tourette syndrome; IV. Syndromes of progressive ataxiasuch as cerebellar degenerations (e.g., cerebellar cortical degenerationand olivopontocerebellar atrophy (OPCA)); and spinocerebellardegeneration (Friedreich's atazia and related disorders); V. Syndrome ofcentral autonomic nervous system failure (Shy-Drager syndrome); VI.Syndromes of muscular weakness and wasting without sensory changes(motoneuron disease such as amyotrophic lateral sclerosis, spinalmuscular atrophy (e.g., infantile spinal muscular atrophy(Werdnig-Hoffman), juvenile spinal muscular atrophy(Wohlfart-Kugelberg-Welander) and other forms of familial spinalmuscular atrophy), primary lateral sclerosis, and hereditary spasticparaplegia; VII. Syndromes combining muscular weakness and wasting withsensory changes (progressive neural muscular atrophy; chronic familialpolyneuropathies) such as peritoneal muscular atrophy(Charcot-Marie-Tooth), hypertrophic interstitial polyneuropathy(Dejerine-Sottas), and miscellaneous forms of chronic progressiveneuropathy; VIII Syndromes of progressive visual loss such as pigmentarydegeneration of the retina (retinitis pigmentosa), and hereditary opticatrophy (Leber's disease). Furthermore, compounds of the invention canbe implicated in chromatin remodeling.

In one aspect, the compounds may also be used in the treatment of adisorder involving, relating to or associated with Src.

Members of the Src-family of tyrosine kinases, in particular, have beenshown to be important in cell signal transduction as it relates toinflammatory response and inflammation-related conditions. Genedisruption studies suggest that inhibition of some members of the srcfamily of kinases would potentially lead to a therapeutic benefit.Src(−/−) mice have abnormalities in bone remodeling or osteopetrosis(Soriano, P. Cell, 1991, 64, 693), suggesting that inhibition of thiskinase might be useful in diseases of bone resorption, such asosteoporosis. Lck(−/−) mice have defects in T cell maturation andactivation (Anderson, S J et al. Adv. Immunol., 1994, 56, 151),suggesting that inhibition of this kinase might be useful in diseases ofT cell mediated inflammation. In addition, human patients have beenidentified with mutations affecting Lck kinase activity (Goldman, F D etal. J. Clin. Invest. 1998, 102, 421). These patients suffer from asevere combined immunodeficiency disorder (SCID).

Src-family kinases are also important for signaling downstream of otherimmune cell receptors. Fyn, like Lck, is involved in TCR signaling in Tcells (Appleby, M W et al. Cell, 1992, 70, 751). Hck and Fgr areinvolved in Fc.gamma. receptor signaling leading to neutrophilactivation (Vicentini, L. et al. J. Immunol. 2002, 168, 6446). Lyn andSrc also participate in Fc.gamma. receptor signaling leading to releaseof histamine and other allergic mediators (Turner, H. and Kinet, J-PNature 1999, 402, B24). These findings suggest that Src family kinaseinhibitors may be useful in treating allergic diseases and asthma.

Src kinases are also activated in tumors including sarcoma, melanoma,breast, and colon cancers suggesting that Src kinase inhibitors may beuseful anti-cancer agents (Abram, C L and Courtneidge, S A Exp. CellRes., 2000, 254, 1). Src kinase inhibitors have also been reported to beeffective in an animal model of cerebral ischemia (R. Paul et al. NatureMedicine, 2001, 7, 222), suggesting that Src kinase inhibitors may beeffective at limiting brain damage following stroke.

Src-family kinases other than Lck, such as Hck and Fgr, are important inthe Fc.gamma. receptor induced respiratory burst of neutrophils as wellas the Fc.gamma. receptor responses of monocytes and macrophages. Thecompounds of the present invention may inhibit the Fey inducedrespiratory burst response in neutrophils, and may also inhibit the Feydependent production of TNFα. The ability to inhibit Fey receptordependent neutrophil, monocyte and macrophage responses would result inadditional anti-inflammatory activity for the present compounds inaddition to their effects on T cells. This activity would be especiallyof value, for example, in the treatment of inflammatory diseases, suchas arthritis or inflammatory bowel disease.

In addition, certain Src family kinases, such as Lyn and Fyn(B), may beimportant in the FCE receptor induced degranulation of mast cells andbasophils that plays an important role in asthma, allergic rhinitis, andother allergic disease. FCE receptors are stimulated by IgE-antigencomplexes. The compounds of the present invention may inhibit the FCEinduced degranulation responses. The ability to inhibit FCE receptordependent mast cell and basophil responses may result in additionalanti-inflammatory activity for the present compounds beyond their effecton T cells.

The invention encompasses pharmaceutical compositions comprisingpharmaceutically acceptable salts of the compounds of the invention asdescribed above. The invention also encompasses pharmaceuticalcompositions comprising hydrates of the compounds of the invention. Theterm “hydrate” includes but is not limited to hemihydrate, monohydrate,dihydrate, trihydrate and the like. The invention further encompassespharmaceutical compositions comprising any solid or liquid physical formof the compound of the invention. For example, the compounds can be in acrystalline form, in amorphous form, and have any particle size. Theparticles may be micronized, or may be agglomerated, particulategranules, powders, oils, oily suspensions or any other form of solid orliquid physical form.

The compounds of the invention, and derivatives, fragments, analogs,homologs pharmaceutically acceptable salts or hydrate thereof can beincorporated into pharmaceutical compositions suitable foradministration, together with a pharmaceutically acceptable carrier orexcipient. Such compositions typically comprise a therapeuticallyeffective amount of any of the compounds above, and a pharmaceuticallyacceptable carrier. Preferably, the effective amount when treatingcancer is an amount effective to selectively induce terminaldifferentiation of suitable neoplastic cells and less than an amountwhich causes toxicity in a patient.

Compounds of the invention may be administered by any suitable means,including, without limitation, parenteral, intravenous, intramuscular,subcutaneous, implantation, oral, sublingual, buccal, nasal, pulmonary,transdermal, topical, vaginal, rectal, and transmucosal administrationsor the like. Topical administration can also involve the use oftransdermal administration such as transdermal patches or iontophoresisdevices. Pharmaceutical preparations include a solid, semisolid orliquid preparation (tablet, pellet, troche, capsule, suppository, cream,ointment, aerosol, powder, liquid, emulsion, suspension, syrup,injection etc.) containing a compound of the invention as an activeingredient, which is suitable for selected mode of administration. Inone embodiment, the pharmaceutical compositions are administered orally,and are thus formulated in a form suitable for oral administration,i.e., as a solid or a liquid preparation. Suitable solid oralformulations include tablets, capsules, pills, granules, pellets,sachets and effervescent, powders, and the like. Suitable liquid oralformulations include solutions, suspensions, dispersions, emulsions,oils and the like. In one embodiment of the present invention, thecomposition is formulated in a capsule. In accordance with thisembodiment, the compositions of the present invention comprise inaddition to the active compound and the inert carrier or diluent, a hardgelatin capsule.

Any inert excipient that is commonly used as a carrier or diluent may beused in the formulations of the present invention, such as for example,a gum, a starch, a sugar, a cellulosic material, an acrylate, ormixtures thereof. A preferred diluent is microcrystalline cellulose. Thecompositions may further comprise a disintegrating agent (e.g.,croscarmellose sodium) and a lubricant (e.g., magnesium stearate), andin addition may comprise one or more additives selected from a binder, abuffer, a protease inhibitor, a surfactant, a solubilizing agent, aplasticizer, an emulsifier, a stabilizing agent, a viscosity increasingagent, a sweetener, a film forming agent, or any combination thereof.Furthermore, the compositions of the present invention may be in theform of controlled release or immediate release formulations.

For liquid formulations, pharmaceutically acceptable carriers may beaqueous or non-aqueous solutions, suspensions, emulsions or oils.Examples of non-aqueous solvents are propylene glycol, polyethyleneglycol, and injectable organic esters such as ethyl oleate. Aqueouscarriers include water, alcoholic/aqueous solutions, emulsions orsuspensions, including saline and buffered media. Examples of oils arethose of petroleum, animal, vegetable, or synthetic origin, for example,peanut oil, soybean oil, mineral oil, olive oil, sunflower oil, andfish-liver oil. Solutions or suspensions can also include the followingcomponents: a sterile diluent such as water for injection, salinesolution, fixed oils, polyethylene glycols, glycerine, propylene glycolor other synthetic solvents; antibacterial agents such as benzyl alcoholor methyl parabens; antioxidants such as ascorbic acid or sodiumbisulfite; chelating agents such as ethylenediaminetetraacetic acid(EDTA); buffers such as acetates, citrates or phosphates, and agents forthe adjustment of tonicity such as sodium chloride or dextrose. The pHcan be adjusted with acids or bases, such as hydrochloric acid or sodiumhydroxide.

In addition, the compositions may further comprise binders (e.g.,acacia, cornstarch, gelatin, carbomer, ethyl cellulose, guar gum,hydroxypropyl cellulose, hydroxypropyl methyl cellulose, povidone),disintegrating agents (e.g., cornstarch, potato starch, alginic acid,silicon dioxide, croscarmellose sodium, crospovidone, guar gum, sodiumstarch glycolate, Primogel), buffers (e.g., tris-HCI., acetate,phosphate) of various pH and ionic strength, additives such as albuminor gelatin to prevent absorption to surfaces, detergents (e.g., Tween20, Tween 80, Pluronic F68, bile acid salts), protease inhibitors,surfactants (e.g., sodium lauryl sulfate), permeation enhancers,solubilizing agents (e.g., glycerol, polyethylene glycerol,cyclodextrins), a glidant (e.g., colloidal silicon dioxide),anti-oxidants (e.g., ascorbic acid, sodium metabisulfite, butylatedhydroxyanisole), stabilizers (e.g., hydroxypropyl cellulose,hyroxypropylmethyl cellulose), viscosity increasing agents (e.g.,carbomer, colloidal silicon dioxide, ethyl cellulose, guar gum),sweeteners (e.g., sucrose, aspartame, citric acid), flavoring agents(e.g., peppermint, methyl salicylate, or orange flavoring),preservatives (e.g., Thimerosal, benzyl alcohol, parabens), lubricants(e.g., stearic acid, magnesium stearate, polyethylene glycol, sodiumlauryl sulfate), flow-aids (e.g., colloidal silicon dioxide),plasticizers (e.g., diethyl phthalate, triethyl citrate), emulsifiers(e.g., carbomer, hydroxypropyl cellulose, sodium lauryl sulfate),polymer coatings (e.g., poloxamers or poloxamines), coating and filmforming agents (e.g., ethyl cellulose, acrylates, polymethacrylates)and/or adjuvants.

In one embodiment, the active compounds are prepared with carriers thatwill protect the compound against rapid elimination from the body, suchas a controlled release formulation, including implants andmicroencapsulated delivery systems. Biodegradable, biocompatiblepolymers can be used, such as ethylene vinyl acetate, polyanhydrides,polyglycolic acid, collagen, polyorthoesters, and polylactic acid.Methods for preparation of such formulations will be apparent to thoseskilled in the art. The materials can also be obtained commercially fromAlza Corporation and Nova Pharmaceuticals, Inc. Liposomal suspensions(including liposomes targeted to infected cells with monoclonalantibodies to viral antigens) can also be used as pharmaceuticallyacceptable carriers. These can be prepared according to methods known tothose skilled in the art, for example, as described in U.S. Pat. No.4,522,811.

It is especially advantageous to formulate oral compositions in dosageunit form for ease of administration and uniformity of dosage. Dosageunit form as used herein refers to physically discrete units suited asunitary dosages for the subject to be treated; each unit containing apredetermined quantity of active compound calculated to produce thedesired therapeutic effect in association with the requiredpharmaceutical carrier. The specification for the dosage unit forms ofthe invention are dictated by and directly dependent on the uniquecharacteristics of the active compound and the particular therapeuticeffect to be achieved, and the limitations inherent in the art ofcompounding such an active compound for the treatment of individuals.

The pharmaceutical compositions can be included in a container, pack, ordispenser together with instructions for administration.

Daily administration may be repeated continuously for a period ofseveral days to several years. Oral treatment may continue for betweenone week and the life of the patient. Preferably the administration maytake place for five consecutive days after which time the patient can beevaluated to determine if further administration is required. Theadministration can be continuous or intermittent, i.e., treatment for anumber of consecutive days followed by a rest period. The compounds ofthe present invention may be administered intravenously on the first dayof treatment, with oral administration on the second day and allconsecutive days thereafter.

The preparation of pharmaceutical compositions that contain an activecomponent is well understood in the art, for example, by mixing,granulating, or tablet-forming processes. The active therapeuticingredient is often mixed with excipients that are pharmaceuticallyacceptable and compatible with the active ingredient. For oraladministration, the active agents are mixed with additives customary forthis purpose, such as vehicles, stabilizers, or inert diluents, andconverted by customary methods into suitable forms for administration,such as tablets, coated tablets, hard or soft gelatin capsules, aqueous,alcoholic or oily solutions and the like as detailed above.

The amount of the compound administered to the patient is less than anamount that would cause toxicity in the patient. In certain embodiments,the amount of the compound that is administered to the patient is lessthan the amount that causes a concentration of the compound in thepatient's plasma to equal or exceed the toxic level of the compound.Preferably, the concentration of the compound in the patient's plasma ismaintained at about 10 nM. In one embodiment, the concentration of thecompound in the patient's plasma is maintained at about 25 nM. In oneembodiment, the concentration of the compound in the patient's plasma ismaintained at about 50 nM. In one embodiment, the concentration of thecompound in the patient's plasma is maintained at about 100 nM. In oneembodiment, the concentration of the compound in the patient's plasma ismaintained at about 500 nM. In one embodiment, the concentration of thecompound in the patient's plasma is maintained at about 1000 nM. In oneembodiment, the concentration of the compound in the patient's plasma ismaintained at about 2500 nM. In one embodiment, the concentration of thecompound in the patient's plasma is maintained at about 5000 nM. Theoptimal amount of the compound that should be administered to thepatient in the practice of the present invention will depend on theparticular compound used and the type of cancer being treated.

DEFINITIONS

Listed below are definitions of various terms used to describe thisinvention. These definitions apply to the terms as they are usedthroughout this specification and claims, unless otherwise limited inspecific instances, either individually or as part of a larger group.

An “aliphatic group” or “aliphatic” is non-aromatic moiety that may besaturated (e.g. single bond) or contain one or more units ofunsaturation, e.g., double and/or triple bonds. An aliphatic group maybe straight chained, branched or cyclic, contain carbon, hydrogen or,optionally, one or more heteroatoms and may be substituted orunsubstituted. An aliphatic group preferably contains between about 1and about 24 atoms, more preferably between about 4 to about 24 atoms,more preferably between about 4-12 atoms, more typically between about 4and about 8 atoms.

The term “acyl” refers to hydrogen, alkyl, partially saturated or fullysaturated cycloalkyl, partially saturated or fully saturatedheterocycle, aryl, and heteroaryl substituted carbonyl groups. Forexample, acyl includes groups such as (C₁-C₆)alkanoyl (e.g., formyl,acetyl, propionyl, butyryl, valeryl, caproyl, t-butylacetyl, etc.),(C₃-C₆)cycloalkylcarbonyl (e.g., cyclopropylcarbonyl,cyclobutylcarbonyl, cyclopentylcarbonyl, cyclohexylcarbonyl, etc.),heterocyclic carbonyl (e.g., pyrrolidinylcarbonyl,pyrrolid-2-one-5-carbonyl, piperidinylcarbonyl, piperazinylcarbonyl,tetrahydrofuranylcarbonyl, etc.), aroyl (e.g., benzoyl) and heteroaroyl(e.g., thiophenyl-2-carbonyl, thiophenyl-3-carbonyl, furanyl-2-carbonyl,furanyl-3-carbonyl, 1H-pyrroyl-2-carbonyl, 1H-pyrroyl-3-carbonyl,benzo[b]thiophenyl-2-carbonyl, etc.). In addition, the alkyl,cycloalkyl, heterocycle, aryl and heteroaryl portion of the acyl groupmay be any one of the groups described in the respective definitions.When indicated as being “optionally substituted”, the acyl group may beunsubstituted or optionally substituted with one or more substituents(typically, one to three substituents) independently selected from thegroup of substituents listed below in the definition for “substituted”or the alkyl, cycloalkyl, heterocycle, aryl and heteroaryl portion ofthe acyl group may be substituted as described above in the preferredand more preferred list of substituents, respectively.

For simplicity, chemical moieties are defined and referred to throughoutcan be univalent chemical moieties (e.g., alkyl, aryl, etc.) ormultivalent moieties under the appropriate structural circumstancesclear to those skilled in the art. For example, an “alkyl” moiety can bereferred to a monovalent radical (e.g. CH₃—CH₂—), or in other instances,a bivalent linking moiety can be “alkyl,” in which case those skilled inthe art will understand the alkyl to be a divalent radical (e.g.,—CH₂—CH₂—), which is equivalent to the term “alkylene.” Similarly, incircumstances in which divalent moieties are required and are stated asbeing “alkoxy”, “alkylamino”, “aryloxy”, “alkylthio”, “aryl”,“heteroaryl”, “heterocyclic”, “alkyl” “alkenyl”, “alkynyl”, “aliphatic”,or “cycloalkyl”, those skilled in the art will understand that the termsalkoxy”, “alkylamino”, “aryloxy”, “alkylthio”, “aryl”, “heteroaryl”,“heterocyclic”, “alkyl”, “alkenyl”, “alkynyl”, “aliphatic”, or“cycloalkyl” refer to the corresponding divalent moiety.

The term “alkyl” embraces linear or branched radicals having one toabout twenty carbon atoms or, preferably, one to about twelve carbonatoms. More preferred alkyl radicals are “lower alkyl” radicals havingone to about ten carbon atoms. Most preferred are lower alkyl radicalshaving one to about eight carbon atoms. Examples of such radicalsinclude methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, pentyl, iso-amyl, hexyl and the like.

The term “alkenyl” embraces linear or branched radicals having at leastone carbon-carbon double bond of two to about twenty carbon atoms or,preferably, two to about twelve carbon atoms. More preferred alkenylradicals are “lower alkenyl” radicals having two to about ten carbonatoms and more preferably about two to about eight carbon atoms.Examples of alkenyl radicals include ethenyl, allyl, propenyl, butenyland 4-methylbutenyl. The terms “alkenyl”, and “lower alkenyl”, embraceradicals having “cis” and “trans” orientations, or alternatively, “E”and “Z” orientations.

The term “alkynyl” embraces linear or branched radicals having at leastone carbon-carbon triple bond of two to about twenty carbon atoms or,preferably, two to about twelve carbon atoms. More preferred alkynylradicals are “lower alkynyl” radicals having two to about ten carbonatoms and more preferably about two to about eight carbon atoms.Examples of alkynyl radicals include propargyl, 1-propynyl, 2-propynyl,1-butyne, 2-butynyl and 1-pentynyl.

The term “cycloalkyl” embraces saturated carbocyclic radicals havingthree to about twelve carbon atoms. The term “cycloalkyl” embracessaturated carbocyclic radicals having three to about twelve carbonatoms. More preferred cycloalkyl radicals are “lower cycloalkyl”radicals having three to about eight carbon atoms. Examples of suchradicals include cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

The term “cycloalkenyl” embraces partially unsaturated carbocyclicradicals having three to twelve carbon atoms. Cycloalkenyl radicals thatare partially unsaturated carbocyclic radicals that contain two doublebonds (that may or may not be conjugated) can be called“cycloalkyldienyl”. More preferred cycloalkenyl radicals are “lowercycloalkenyl” radicals having four to about eight carbon atoms. Examplesof such radicals include cyclobutenyl, cyclopentenyl and cyclohexenyl.

The terms “alkoxy” embrace linear or branched oxy-containing radicalseach having alkyl portions of one to about twenty carbon atoms or,preferably, one to about twelve carbon atoms. More preferred alkoxyradicals are “lower alkoxy” radicals having one to about ten carbonatoms and more preferably having one to about eight carbon atoms.Examples of such radicals include methoxy, ethoxy, propoxy, butoxy andtert-butoxy.

The term “alkoxyalkyl” embraces alkyl radicals having one or more alkoxyradicals attached to the alkyl radical, that is, to form monoalkoxyalkyland dialkoxyalkyl radicals.

The term “aryl”, alone or in combination, means a carbocyclic aromaticsystem containing one, two or three rings wherein such rings may beattached together in a pendent manner or may be fused. The term “aryl”embraces aromatic radicals such as phenyl, naphthyl, tetrahydronaphthyl,indane and biphenyl.

The term “carbonyl”, whether used alone or with other terms, such as“alkoxycarbonyl”, denotes (C═O).

The term “carbanoyl”, whether used alone or with other terms, such as“arylcarbanoylyalkyl”, denotes C(O)NH.

The terms “heterocyclyl”, “heterocycle” “heterocyclic” or “heterocyclo”embrace saturated, partially unsaturated and unsaturatedheteroatom-containing ring-shaped radicals, which can also be called“heterocyclyl”, “heterocycloalkenyl” and “heteroaryl” correspondingly,where the heteroatoms may be selected from nitrogen, sulfur and oxygen.Examples of saturated heterocyclyl radicals include saturated 3 to6-membered heteromonocyclic group containing 1 to 4 nitrogen atoms (e.g.pyrrolidinyl, imidazolidinyl, piperidino, piperazinyl, etc.); saturated3 to 6-membered heteromonocyclic group containing 1 to 2 oxygen atomsand 1 to 3 nitrogen atoms (e.g. morpholinyl, etc.); saturated 3 to6-membered heteromonocyclic group containing 1 to 2 sulfur atoms and 1to 3 nitrogen atoms (e.g., thiazolidinyl, etc.). Examples of partiallyunsaturated heterocyclyl radicals include dihydrothiophene,dihydropyran, dihydrofuran and dihydrothiazole. Heterocyclyl radicalsmay include a pentavalent nitrogen, such as in tetrazolium andpyridinium radicals. The term “heterocycle” also embraces radicals whereheterocyclyl radicals are fused with aryl or cycloalkyl radicals.Examples of such fused bicyclic radicals include benzofuran,benzothiophene, and the like.

The term “heteroaryl” embraces unsaturated heterocyclyl radicals.Examples of heteroaryl radicals include unsaturated 3 to 6 memberedheteromonocyclic group containing 1 to 4 nitrogen atoms, for example,pyrrolyl, pyrrolinyl, imidazolyl, pyrazolyl, pyridyl, pyrimidyl,pyrazinyl, pyridazinyl, triazolyl (e.g., 4H-1,2,4-triazolyl,1H-1,2,3-triazolyl, 2H-1,2,3-triazolyl, etc.), tetrazolyl (e.g.1H-tetrazolyl, 2H-tetrazolyl, etc.), etc.; unsaturated condensedheterocyclyl group containing 1 to 5 nitrogen atoms, for example,indolyl, isoindolyl, indolizinyl, benzimidazolyl, quinolyl, isoquinolyl,indazolyl, benzotriazolyl, tetrazolopyridazinyl (e.g.,tetrazolo[1,5-b]pyridazinyl, etc.), etc.; unsaturated 3 to 6-memberedheteromonocyclic group containing an oxygen atom, for example, pyranyl,furyl, etc.; unsaturated 3 to 6-membered heteromonocyclic groupcontaining a sulfur atom, for example, thienyl, etc.; unsaturated 3- to6-membered heteromonocyclic group containing 1 to 2 oxygen atoms and 1to 3 nitrogen atoms, for example, oxazolyl, isoxazolyl, oxadiazolyl(e.g., 1,2,4-oxadiazolyl, 1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl, etc.)etc.; unsaturated condensed heterocyclyl group containing 1 to 2 oxygenatoms and 1 to 3 nitrogen atoms (e.g. benzoxazolyl, benzoxadiazolyl,etc.); unsaturated 3 to 6-membered heteromonocyclic group containing 1to 2 sulfur atoms and 1 to 3 nitrogen atoms, for example, thiazolyl,thiadiazolyl (e.g., 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl,1,2,5-thiadiazolyl, etc.) etc.; unsaturated condensed heterocyclyl groupcontaining 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms (e.g.,benzothiazolyl, benzothiadiazolyl, etc.) and the like.

The term “heterocycloalkyl” embraces heterocyclo-substituted alkylradicals. More preferred heterocycloalkyl radicals are “lowerheterocycloalkyl” radicals having one to six carbon atoms andheterocyclo radicals.

The term “alkylthio” embraces radicals containing a linear or branchedalkyl radical, of one to about ten carbon atoms attached to a divalentsulfur atom. Preferred alkylthio radicals have alkyl radicals of one toabout twenty carbon atoms or, preferably, one to about twelve carbonatoms. More preferred alkylthio radicals have alkyl radicals are “loweralkylthio” radicals having one to about ten carbon atoms. Most preferredare alkylthio radicals having lower alkyl radicals of one to about eightcarbon atoms. Examples of such lower alkylthio radicals are methylthio,ethylthio, propylthio, butylthio and hexylthio.

The terms “aralkyl” or “arylalkyl” embrace aryl-substituted alkylradicals such as benzyl, diphenylmethyl, triphenylmethyl, phenylethyl,and diphenylethyl.

The term “aryloxy” embraces aryl radicals attached through an oxygenatom to other radicals.

The terms “aralkoxy” or “arylalkoxy” embrace aralkyl radicals attachedthrough an oxygen atom to other radicals.

The term “aminoalkyl” embraces alkyl radicals substituted with aminoradicals. Preferred aminoalkyl radicals have alkyl radicals having aboutone to about twenty carbon atoms or, preferably, one to about twelvecarbon atoms. More preferred aminoalkyl radicals are “lower aminoalkyl”that have alkyl radicals having one to about ten carbon atoms. Mostpreferred are aminoalkyl radicals having lower alkyl radicals having oneto eight carbon atoms. Examples of such radicals include aminomethyl,aminoethyl, and the like.

The term “alkylamino” denotes amino groups which are substituted withone or two alkyl radicals. Preferred alkylamino radicals have alkylradicals having about one to about twenty carbon atoms or, preferably,one to about twelve carbon atoms. More preferred alkylamino radicals are“lower alkylamino” that have alkyl radicals having one to about tencarbon atoms. Most preferred are alkylamino radicals having lower alkylradicals having one to about eight carbon atoms. Suitable loweralkylamino may be monosubstituted N-alkylamino or disubstitutedN,N-alkylamino, such as N-methylamino, N-ethylamino, N,N-dimethylamino,N,N-diethylamino or the like.

The term “linker” means an organic moiety that connects two parts of acompound. Linkers typically comprise a direct bond or an atom such asoxygen or sulfur, a unit such as NR₈, C(O), C(O)NH, SO, SO₂, SO₂NH or achain of atoms, such as substituted or unsubstituted alkyl, substitutedor unsubstituted alkenyl, substituted or unsubstituted alkynyl,arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl, heteroarylalkenyl,heteroarylalkynyl, heterocyclylalkyl, heterocyclylalkenyl,heterocyclylalkynyl, aryl, heteroaryl, heterocyclyl, cycloalkyl,cycloalkenyl, alkylarylalkyl, alkylarylalkenyl, alkylarylalkynyl,alkenylarylalkyl, alkenylarylalkenyl, alkenylarylalkynyl,alkynylarylalkyl, alkynylarylalkenyl, alkynylarylalkynyl,alkylheteroarylalkyl, alkylheteroarylalkenyl, alkylheteroarylalkynyl,alkenylheteroarylalkyl, alkenylheteroarylalkenyl,alkenylheteroarylalkynyl, alkynylheteroarylalkyl,alkynylheteroarylalkenyl, alkynylheteroarylalkynyl,alkylheterocyclylalkyl, alkylheterocyclylalkenyl,alkylhererocyclylalkynyl, alkenylheterocyclylalkyl,alkenylheterocyclylalkenyl, alkenylheterocyclylalkynyl,alkynylheterocyclylalkyl, alkynylheterocyclylalkenyl,alkynylheterocyclylalkynyl, alkylaryl, alkenylaryl, alkynylaryl,alkylheteroaryl, alkenylheteroaryl, alkynylhereroaryl, which one or moremethylenes can be interrupted or terminated by O, S, S(O), SO₂, N(R₈),C(O), substituted or unsubstituted aryl, substituted or unsubstitutedheteroaryl, substituted or unsubstituted heterocyclic; where R₈ ishydrogen, acyl, aliphatic or substituted aliphatic. In one embodiment,the linker B is between 1-24 atoms, preferably 4-24 atoms, preferably4-18 atoms, more preferably 4-12 atoms, and most preferably about 4-10atoms.

The term “substituted” refers to the replacement of one or more hydrogenradicals in a given structure with the radical of a specifiedsubstituent including, but not limited to, halo, alkyl, alkenyl,alkynyl, aryl, heterocyclyl, thiol, alkylthio, arylthio, alkylthioalkyl,arylthioalkyl, alkylsulfonyl, alkylsulfonylalkyl, arylsulfonylalkyl,alkoxy, aryloxy, aralkoxy, aminocarbonyl, alkylaminocarbonyl,arylaminocarbonyl, alkoxycarbonyl, aryloxycarbonyl, haloalkyl, amino,trifluoromethyl, cyano, nitro, alkylamino, arylamino, alkylaminoalkyl,arylaminoalkyl, aminoalkylamino, hydroxy, alkoxyalkyl, carboxyalkyl,alkoxycarbonylalkyl, aminocarbonylalkyl, acyl, aralkoxycarbonyl,carboxylic acid, sulfonic acid, sulfonyl, phosphonic acid, aryl,heteroaryl, heterocyclic, and aliphatic. It is understood that thesubstituents listed above may be further substituted.

The term “halogen” or “halo” as used herein, refers to an atom selectedfrom fluorine, chlorine, bromine and iodine.

As used herein, the term “aberrant proliferation” refers to abnormalcell growth.

The phrase “adjunctive therapy” encompasses treatment of a subject withagents that reduce or avoid side effects associated with the combinationtherapy of the present invention, including, but not limited to, thoseagents, for example, that reduce the toxic effect of anticancer drugs,e.g., bone resorption inhibitors, cardioprotective agents; prevent orreduce the incidence of nausea and vomiting associated withchemotherapy, radiotherapy or operation; or reduce the incidence ofinfection associated with the administration of myelosuppressiveanticancer drugs.

The term “angiogenesis,” as used herein, refers to the formation ofblood vessels. Specifically, angiogenesis is a multi-step process inwhich endothelial cells focally degrade and invade through their ownbasement membrane, migrate through interstitial stroma toward anangiogenic stimulus, proliferate proximal to the migrating tip, organizeinto blood vessels, and reattach to newly synthesized basement membrane(see Folkman et al., Adv. Cancer Res., Vol. 43, pp. 175-203 (1985)).Anti-angiogenic agents interfere with this process. Examples of agentsthat interfere with several of these steps include thrombospondin-1,angiostatin, endostatin, interferon alpha and compounds such as matrixmetalloproteinase (MMP) inhibitors that block the actions of enzymesthat clear and create paths for newly forming blood vessels to follow;compounds, such as .alpha.v.beta.3 inhibitors, that interfere withmolecules that blood vessel cells use to bridge between a parent bloodvessel and a tumor; agents, such as specific COX-2 inhibitors, thatprevent the growth of cells that form new blood vessels; andprotein-based compounds that simultaneously interfere with several ofthese targets.

The term “apoptosis” as used herein refers to programmed cell death assignaled by the nuclei in normally functioning human and animal cellswhen age or state of cell health and condition dictates. An “apoptosisinducing agent” triggers the process of programmed cell death.

The term “cancer” as used herein denotes a class of diseases ordisorders characterized by uncontrolled division of cells and theability of these cells to invade other tissues, either by direct growthinto adjacent tissue through invasion or by implantation into distantsites by metastasis.

The term “compound” is defined herein to include pharmaceuticallyacceptable salts, solvates, hydrates, polymorphs, enantiomers,diastereoisomers, racemates and the like of the compounds having aformula as set forth herein.

The term “devices” refers to any appliance, usually mechanical orelectrical, designed to perform a particular function.

As used herein, the term “dysplasia” refers to abnormal cell growth, andtypically refers to the earliest form of pre-cancerous lesionrecognizable in a biopsy by a pathologist.

As used herein, the term “effective amount of the subject compounds,”with respect to the subject method of treatment, refers to an amount ofthe subject compound which, when delivered as part of desired doseregimen, brings about, e.g. a change in the rate of cell proliferationand/or state of differentiation and/or rate of survival of a cell toclinically acceptable standards. This amount may further relieve to someextent one or more of the symptoms of a neoplasia disorder, including,but is not limited to: 1) reduction in the number of cancer cells; 2)reduction in tumor size; 3) inhibition (i.e., slowing to some extent,preferably stopping) of cancer cell infiltration into peripheral organs;4) inhibition (i.e., slowing to some extent, preferably stopping) oftumor metastasis; 5) inhibition, to some extent, of tumor growth; 6)relieving or reducing to some extent one or more of the symptomsassociated with the disorder; and/or 7) relieving or reducing the sideeffects associated with the administration of anticancer agents.

The term “hyperplasia,” as used herein, refers to excessive celldivision or growth.

The phrase an “immunotherapeutic agent” refers to agents used totransfer the immunity of an immune donor, e.g., another person or ananimal, to a host by inoculation. The term embraces the use of serum orgamma globulin containing performed antibodies produced by anotherindividual or an animal; nonspecific systemic stimulation; adjuvants;active specific immunotherapy; and adoptive immunotherapy. Adoptiveimmunotherapy refers to the treatment of a disease by therapy or agentsthat include host inoculation of sensitized lymphocytes, transferfactor, immune RNA, or antibodies in serum or gamma globulin.

The term “inhibition,” in the context of neoplasia, tumor growth ortumor cell growth, may be assessed by delayed appearance of primary orsecondary tumors, slowed development of primary or secondary tumors,decreased occurrence of primary or secondary tumors, slowed or decreasedseverity of secondary effects of disease, arrested tumor growth andregression of tumors, among others. In the extreme, complete inhibition,is referred to herein as prevention or chemoprevention.

The term “metastasis,” as used herein, refers to the migration of cancercells from the original tumor site through the blood and lymph vesselsto produce cancers in other tissues. Metastasis also is the term usedfor a secondary cancer growing at a distant site.

The term “neoplasm,” as used herein, refers to an abnormal mass oftissue that results from excessive cell division. Neoplasms may bebenign (not cancerous), or malignant (cancerous) and may also be calleda tumor. The term “neoplasia” is the pathological process that resultsin tumor formation.

As used herein, the term “pre-cancerous” refers to a condition that isnot malignant, but is likely to become malignant if left untreated.

The term “proliferation” refers to cells undergoing mitosis.

The phrase a “radiotherapeutic agent” refers to the use ofelectromagnetic or particulate radiation in the treatment of neoplasia.

The term “recurrence” as used herein refers to the return of cancerafter a period of remission. This may be due to incomplete removal ofcells from the initial cancer and may occur locally (the same site ofinitial cancer), regionally (in vicinity of initial cancer, possibly inthe lymph nodes or tissue), and/or distally as a result of metastasis.

The term “treatment” refers to any process, action, application,therapy, or the like, wherein a mammal, including a human being, issubject to medical aid with the object of improving the mammal'scondition, directly or indirectly.

The term “vaccine” includes agents that induce the patient's immunesystem to mount an immune response against the tumor by attacking cellsthat express tumor associated antigens (TAAs).

As used herein, the term “effective amount of the subject compounds,”with respect to the subject method of treatment, refers to an amount ofthe subject compound which, when delivered as part of desired doseregimen, brings about, e.g. a change in the rate of cell proliferationand/or state of differentiation and/or rate of survival of a cell toclinically acceptable standards. This amount may further relieve to someextent one or more of the symptoms of a neoplasia disorder, including,but is not limited to: 1) reduction in the number of cancer cells; 2)reduction in tumor size; 3) inhibition (i.e., slowing to some extent,preferably stopping) of cancer cell infiltration into peripheral organs;4) inhibition (i.e., slowing to some extent, preferably stopping) oftumor metastasis; 5) inhibition, to some extent, of tumor growth; 6)relieving or reducing to some extent one or more of the symptomsassociated with the disorder; and/or 7) relieving or reducing the sideeffects associated with the administration of anticancer agents.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge, etal. describes pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 66: 1-19 (1977). The salts can be prepared insitu during the final isolation and purification of the compounds of theinvention, or separately by reacting the free base function with asuitable organic acid or inorganic acid. Examples of pharmaceuticallyacceptable nontoxic acid addition salts include, but are not limited to,salts of an amino group formed with inorganic acids such as hydrochloricacid, hydrobromic acid, phosphoric acid, sulfuric acid and perchloricacid or with organic acids such as acetic acid, maleic acid, tartaricacid, citric acid, succinic acid lactobionic acid or malonic acid or byusing other methods used in the art such as ion exchange. Otherpharmaceutically acceptable salts include, but are not limited to,adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,palmoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Representative alkali or alkaline earth metal salts includesodium, lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, alkyl having from 1 to 6 carbon atoms, sulfonate and arylsulfonate.

As used herein, the term “pharmaceutically acceptable ester” refers toesters which hydrolyze in vivo and include those that break down readilyin the human body to leave the parent compound or a salt thereof.Suitable ester groups include, for example, those derived frompharmaceutically acceptable aliphatic carboxylic acids, particularlyalkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which eachalkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.Examples of particular esters include, but are not limited to, formates,acetates, propionates, butyrates, acrylates and ethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds of the present invention which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and lower animals with undue toxicity,irritation, allergic response, and the like, commensurate with areasonable benefit/risk ratio, and effective for their intended use, aswell as the zwitterionic forms, where possible, of the compounds of thepresent invention. “Prodrug”, as used herein means a compound which isconvertible in vivo by metabolic means (e.g. by hydrolysis) to acompound of the invention. Various forms of prodrugs are known in theart, for example, as discussed in Bundgaard, (ed.), Design of Prodrugs,Elsevier (1985); Widder, et al. (ed.), Methods in Enzymology, vol. 4,Academic Press (1985); Krogsgaard-Larsen, et al., (ed). “Design andApplication of Prodrugs, Textbook of Drug Design and Development,Chapter 5, 113-191 (1991); Bundgaard, et al., Journal of Drug DeliverReviews, 8:1-38 (1992); Bundgaard, J. of Pharmaceutical Sciences, 77:285et seq. (1988); Higuchi and Stella (eds.) Prodrugs as Novel DrugDelivery Systems, American Chemical Society (1975); and Bernard Testa &Joachim Mayer, “Hydrolysis In Drug And Prodrug Metabolism: Chemistry,Biochemistry And Enzymology,” John Wiley and Sons, Ltd. (2002).

As used herein, “pharmaceutically acceptable carrier” is intended toinclude any and all solvents, dispersion media, coatings, antibacterialand antifungal agents, isotonic and absorption delaying agents, and thelike, compatible with pharmaceutical administration, such as sterilepyrogen-free water. Suitable carriers are described in the most recentedition of Remington's Pharmaceutical Sciences, a standard referencetext in the field, which is incorporated herein by reference. Preferredexamples of such carriers or diluents include, but are not limited to,water, saline, finger's solutions, dextrose solution, and 5% human serumalbumin. Liposomes and non-aqueous vehicles such as fixed oils may alsobe used. The use of such media and agents for pharmaceutically activesubstances is well known in the art. Except insofar as any conventionalmedia or agent is incompatible with the active compound, use thereof inthe compositions is contemplated. Supplementary active compounds canalso be incorporated into the compositions.

As used herein, the term “pre-cancerous” refers to a condition that isnot malignant, but is likely to become malignant if left untreated.

The term “subject” as used herein refers to an animal. Preferably theanimal is a mammal. More preferably the mammal is a human. A subjectalso refers to, for example, dogs, cats, horses, cows, pigs, guineapigs, fish, birds and the like.

The compounds of this invention may be modified by appending appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and may include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of excretion.

The synthesized compounds can be separated from a reaction mixture andfurther purified by a method such as column chromatography, highpressure liquid chromatography, or recrystallization. As can beappreciated by the skilled artisan, further methods of synthesizing thecompounds of the formulae herein will be evident to those of ordinaryskill in the art. Additionally, the various synthetic steps may beperformed in an alternate sequence or order to give the desiredcompounds. Synthetic chemistry transformations and protecting groupmethodologies (protection and deprotection) useful in synthesizing thecompounds described herein are known in the art and include, forexample, those such as described in R. Larock, Comprehensive OrganicTransformations, VCH Publishers (1989); T. W. Greene and P. G. M. Wuts,Protective Groups in Organic Synthesis, 2d. Ed., John Wiley and Sons(1991); L. Fieser and M. Fieser, Fieser and Fieser's Reagents forOrganic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons(1995), and subsequent editions thereof.

The compounds described herein contain one or more asymmetric centersand thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms that may be defined, in terms of absolutestereochemistry, as (R)- or (S)-, or as (D)- or (L)- for amino acids.The present invention is meant to include all such possible isomers, aswell as their racemic and optically pure forms. Optical isomers may beprepared from their respective optically active precursors by theprocedures described above, or by resolving the racemic mixtures. Theresolution can be carried out in the presence of a resolving agent, bychromatography or by repeated crystallization or by some combination ofthese techniques which are known to those skilled in the art. Furtherdetails regarding resolutions can be found in Jacques, et al.,Enantiomers, Racemates, and Resolutions (John Wiley & Sons, 1981). Whenthe compounds described herein contain olefinic double bonds, otherunsaturation, or other centers of geometric asymmetry, and unlessspecified otherwise, it is intended that the compounds include both Eand Z geometric isomers or cis- and trans-isomers. Likewise, alltautomeric forms are also intended to be included. The configuration ofany carbon-carbon double bond appearing herein is selected forconvenience only and is not intended to designate a particularconfiguration unless the text so states; thus a carbon-carbon doublebond or carbon-heteroatom double bond depicted arbitrarily herein astrans may be cis, trans, or a mixture of the two in any proportion.

Pharmaceutical Compositions

The pharmaceutical compositions of the present invention comprise atherapeutically effective amount of a compound of the present inventionformulated together with one or more pharmaceutically acceptablecarriers or excipients.

As used herein, the term “pharmaceutically acceptable carrier orexcipient” means a non-toxic, inert solid, semi-solid or liquid filler,diluent, encapsulating material or formulation auxiliary of any type.Some examples of materials which can serve as pharmaceuticallyacceptable carriers are sugars such as lactose, glucose and sucrose;cyclodextrins such as alpha-(α), beta-(B) and gamma-(γ) cyclodextrins;starches such as corn starch and potato starch; cellulose and itsderivatives such as sodium carboxymethyl cellulose, ethyl cellulose andcellulose acetate; powdered tragacanth; malt; gelatin; talc; excipientssuch as cocoa butter and suppository waxes; oils such as peanut oil,cottonseed oil, safflower oil, sesame oil, olive oil, corn oil andsoybean oil; glycols such as propylene glycol; esters such as ethyloleate and ethyl laurate; agar; buffering agents such as magnesiumhydroxide and aluminum hydroxide; alginic acid; pyrogen-free water;isotonic saline; Ringer's solution; ethyl alcohol, and phosphate buffersolutions, as well as other non-toxic compatible lubricants such assodium lauryl sulfate and magnesium stearate, as well as coloringagents, releasing agents, coating agents, sweetening, flavoring andperfuming agents, preservatives and antioxidants can also be present inthe composition, according to the judgment of the formulator.

The pharmaceutical compositions of this invention may be administeredorally, parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir, preferably by oraladministration or administration by injection. The pharmaceuticalcompositions of this invention may contain any conventional non-toxicpharmaceutically-acceptable carriers, adjuvants or vehicles. In somecases, the pH of the formulation may be adjusted with pharmaceuticallyacceptable acids, bases or buffers to enhance the stability of theformulated compound or its delivery form. The term parenteral as usedherein includes subcutaneous, intracutaneous, intravenous,intramuscular, intra-articular, intraarterial, intrasynovial,intrasternal, intrathecal, intralesional and intracranial injection orinfusion techniques.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils (in particular, cottonseed, groundnut,corn, germ, olive, castor, and sesame oils), glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitan, and mixtures thereof. Besides inert diluents, the oralcompositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions, may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolution,which, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform is accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations are also prepared by entrapping the drug in liposomes ormicroemulsions that are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or: a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, eye ointments, powders and solutionsare also contemplated as being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Powders and sprays can contain, in addition to the compounds of thisinvention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel.

For pulmonary delivery, a therapeutic composition of the invention isformulated and administered to the patient in solid or liquidparticulate form by direct administration e.g., inhalation into therespiratory system. Solid or liquid particulate forms of the activecompound prepared for practicing the present invention include particlesof respirable size: that is, particles of a size sufficiently small topass through the mouth and larynx upon inhalation and into the bronchiand alveoli of the lungs. Delivery of aerosolized therapeutics,particularly aerosolized antibiotics, is known in the art (see, forexample U.S. Pat. No. 5,767,068 to VanDevanter et al., U.S. Pat. No.5,508,269 to Smith et al., and WO 98/43650 by Montgomery, all of whichare incorporated herein by reference). A discussion of pulmonarydelivery of antibiotics is also found in U.S. Pat. No. 6,014,969,incorporated herein by reference.

By a “therapeutically effective amount” of a compound of the inventionis meant an amount of the compound which confers a therapeutic effect onthe treated subject, at a reasonable benefit/risk ratio applicable toany medical treatment. The therapeutic effect may be objective (i.e.,measurable by some test or marker) or subjective (i.e., subject gives anindication of or feels an effect). An effective amount of the compounddescribed above may range from about 0.1 mg/Kg to about 500 mg/Kg,preferably from about 1 to about 50 mg/Kg. Effective doses will alsovary depending on route of administration, as well as the possibility ofco-usage with other agents. It will be understood, however, that thetotal daily usage of the compounds and compositions of the presentinvention will be decided by the attending physician within the scope ofsound medical judgment. The specific therapeutically effective doselevel for any particular patient will depend upon a variety of factorsincluding the disorder being treated and the severity of the disorder;the activity of the specific compound employed; the specific compositionemployed; the age, body weight, general health, sex and diet of thepatient; the time of administration, route of administration, and rateof excretion of the specific compound employed; the duration of thetreatment; drugs used in combination or contemporaneously with thespecific compound employed; and like factors well known in the medicalarts.

The total daily dose of the compounds of this invention administered toa human or other animal in single or in divided doses can be in amounts,for example, from 0.01 to 50 mg/kg body weight or more usually from 0.1to 25 mg/kg body weight. Single dose compositions may contain suchamounts or submultiples thereof to make up the daily dose. In general,treatment regimens according to the present invention compriseadministration to a patient in need of such treatment from about 10 mgto about 1000 mg of the compound(s) of this invention per day in singleor multiple doses.

The compounds of the formulae described herein can, for example, beadministered by injection, intravenously, intraarterially, subdermally,intraperitoneally, intramuscularly, or subcutaneously; or orally,buccally, nasally, transmucosally, topically, in an ophthalmicpreparation, or by inhalation, with a dosage ranging from about 0.1 toabout 500 mg/kg of body weight, alternatively dosages between 1 mg and1000 mg/dose, every 4 to 120 hours, or according to the requirements ofthe particular drug. The methods herein contemplate administration of aneffective amount of compound or compound composition to achieve thedesired or stated effect. Typically, the pharmaceutical compositions ofthis invention will be administered from about 1 to about 6 times perday or alternatively, as a continuous infusion. Such administration canbe used as a chronic or acute therapy. The amount of active ingredientthat may be combined with pharmaceutically excipients or carriers toproduce a single dosage form will vary depending upon the host treatedand the particular mode of administration. A typical preparation willcontain from about 5% to about 95% active compound (w/w). Alternatively,such preparations may contain from about 20% to about 80% activecompound.

Lower or higher doses than those recited above may be required. Specificdosage and treatment regimens for any particular patient will dependupon a variety of factors, including the activity of the specificcompound employed, the age, body weight, general health status, sex,diet, time of administration, rate of excretion, drug combination, theseverity and course of the disease, condition or symptoms, the patient'sdisposition to the disease, condition or symptoms, and the judgment ofthe treating physician.

Upon improvement of a patient's condition, a maintenance dose of acompound, composition or combination of this invention may beadministered, if necessary. Subsequently, the dosage or frequency ofadministration, or both, may be reduced, as a function of the symptoms,to a level at which the improved condition is retained when the symptomshave been alleviated to the desired level. Patients may, however,require intermittent treatment on a long-term basis upon any recurrenceof disease symptoms.

Synthetic Methods

The compounds of the invention may be prepared by any process known tobe applicable to the preparation of chemically-related compounds.Suitable processes for making certain intermediates include, forexample, those illustrated references such as, J. Med. Chem. 2004, 47,6658-6661 and J. Org. Chem. 1952, 1320. Necessary starting materials maybe obtained by standard procedures of organic chemistry. The preparationof such starting materials is described within the accompanyingnon-limiting Examples. Alternatively necessary starting materials areobtainable by analogous procedures to those illustrated which are withinthe ordinary skill of a chemist.

The compounds and processes of the present invention will be betterunderstood in connection with the following representative syntheticscheme that illustrate the methods by which the compounds of theinvention may be prepared, which are intended as an illustration onlyand not limiting of the scope of the invention.

EXAMPLES

The compounds and processes of the present invention will be betterunderstood in connection with the following examples, which are intendedas an illustration only and not limiting of the scope of the invention.Various changes and modifications to the disclosed embodiments will beapparent to those skilled in the art and such changes and modificationsincluding, without limitation, those relating to the chemicalstructures, substituents, derivatives, formulations and/or methods ofthe invention may be made without departing from the spirit of theinvention and the scope of the appended claims.

Example 1 Preparation ofN-(2-chloro-6-methylphenyl)-2-(6-(4-(2-(hydroxyamino)-2-oxoethyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 1) Step 1a. 2-Chlorothiazole (Compound 102)

A solution of 2-aminothiazole (101) (20.0 g, 200.0 mmol) in saturatedaqueous NaCl (20 mL) and HCl (60 mL) was maintained in aroom-temperature bath. It was then treated with NaNO₂ (250 mmol) in H₂O(50 mL) and concentrated HCl (20 mL) dropwise simultaneously. Thereaction was stirred at room temperature for 1 hour, extracted withether and concentrated at 1 atm. The product was obtained by distilledunder vacuum to give 102 as a pale yellow liquid (10.7 g, 45%): ¹H NMR(CDCl₃) δ 7.24 (d, J=3.6 Hz, 1H), 7.57 (d, J=3.3 Hz, 1H).

Step 1b. 2-Chloro-N-(2-chloro-6-methylphenyl)thiazole-5-carboxamide(Compound 103)

A solution of 2-chlorothiazole (102) (480 mg, 4.0 mmol) in THF (10 mL)was cooled to −78° C. and treated dropwise with 2.5 M n-butyllithium inhexanes (1.68 mL, 4.2 mmol) over a period of 20 minutes while keepingthe temperature below −75° C. After the addition was complete, themixture was stirred at −78° C. for 15 minutes and then treated with asolution of 2-chloro-6-methylphenylisocyanate (4.4 mmol) in THF (5 mL).The mixture was stirred at −78° C. for 2 hours, quenched with saturatedaqueous NH₄Cl, warmed to room temperature and partitioned between EtOAcand H₂O. The EtOAc phase was separated, washed with brine, dried(Na₂SO₄) and concentrated under vacuum to afford a yellow solid. Thecrude product was purified by column chromatography to give compound 103as a pale yellow solid (0.95 g, 83%). LCMS: 286 [M+1]⁺. ¹H NMR(DMSO-d₆): δ 2.22 (s, 3H), 7.29 (m, 2H), 7.41 (dd, J=6.3, J=3.0 Hz, 1H),8.45 (s, 1H), 10.40 (s, 1H).

Step 1c.2-Chloro-N-(2-chloro-6-methylphenyl)-N-(4-methoxybenzyl)thiazole-5-carboxamide(Compound 104)

A solution of 2-chloro-N-(2-chloro-6-methylphenyl)thiazole (103) (0.57g, 2.0 mmol) in DMF (5 mL) was treated with 60% NaH (2.4 mmol) andstirred at room temperature for 30 minutes. The mixture was treated with4-methoxybenzyl chloride (0.38 g, 2.4 mmol) and tetrabutylammoniumiodide (0.15 mg, 0.40 mmol), and then stirred at room temperature for 16h. The mixture was partitioned between H₂O and EtOAc and then the EtOAcphase was separated, washed with brine, dried (Na₂SO₄) and concentratedunder vacuum. The crude product was purified by column chromatography togive compound 104 as a yellow solid (0.50 g, 62%). LCMS: 429 [M+Na]⁺. ¹HNMR (DMSO-d₆): δ 1.73 (s, 3H), 3.60 (s, 3H), 4.48 (d, J=13.8 Hz, 1H),5.09 (d, J=14.1 Hz, 1H), 6.79 (d, J=8.4 Hz, 2H), 7.11 (d, J=8.7 Hz, 2H),7.29 (d, J=6.6 Hz, 1H), 7.44 (m, 3H).

Step 1d. 6-Chloro-2-methylpyrimidin-4-amine (Compound 106)

A solution of 4,6-dichloro-2-methylpyrimidin (105) (20.0 g, 120 mmol)was placed in a tube with ammonium hydroxide (50 mL). The tube wassealed and heated at 125-128° C. for 10 hours. After cooling, the tubewas opened and the reaction mixture (coarse, white crystals) wasfiltrated, giving the product compound 106 as a white solid (12.4 g,70%). LCMS: 144 [M+1]⁺ ¹H NMR (DMSO-d₆): δ 2.27 (s, 3H), 6.24 (s, 1H),7.08 (s, 2H).

Step 1e.2-((6-Chloro-2-methylpyrimidin-4-yl)methyl)-N-(2-chloro-6-methylphenyl)-N-(4-methoxybenzyl)thiazole-5-carboxamide(Compound 107)

4-Amino-6-chloro-2-methylpyrimidine (106) (14.0 mg, 0.10 mmol) was addedin portions to a suspension of NaH (60% dispersion, 0.30 mmol) in THF(30 mL) at 0° C. for 30 minutes and then treated with compound 104 (41.0mg, 0.10 mmol) in portions. The resulting mixture was at reflux for 4hours, cooled to room temperature and diluted with H₂O (10 mL). Themixture was acidified with 1 N HCl (5 mL) and extracted with EtOAc (3×10mL). The organic layer was dried (Na₂SO₄) and evaporated. The crudeproduct was purified by column chromatography to give compound 107 as apale yellow solid (41 mg, 80%): ¹H NMR (DMSO-d₆): δ 1.72 (s, 3H), 2.45(s, 3H), 3.71 (s, 3H), 4.40 (d, J=14.1 Hz, 1H) 5.19 (d, J=13.8 Hz, 1H),6.81 (m, 3H), 7.14 (d, J=8.4 Hz, 2H), 7.29 (d, J=7.5 Hz, 1H), 7.43 (t,J=7.8, Hz, 1H), 7.47 (s, 1H), 7.53 (d, J=6.9 Hz, 1H), 12.07 (ds, 1H).

Step 1f.2-(6-Chloro-2-methylpyrimidin-4-ylamino)-N-(2-chloro-6-methylphenyl)thiazole-5-carboxamide(Compound 108)

A solution of compound 107 (10.0 g, 19.5 mmol) dissolved in 50% TFA inCH₂Cl₂ (50 mL) was treated with triflic acid (10.0 g, 67.5 mmol). Thereaction mixture was stirred at room temperature for 24 hours. Themixture was poured into crushed ice (150 g). The resulting solid wascollected by filtration to obtain compound 108 as a yellow solid (5.6 g,87%). ¹H NMR (DMSO-d₆): δ 2.21 (s, 3H), 2.39 (s, 3H), 6.07 (m, 1H), 7.26(m, 2H), 7.37 (d, J=6.6 Hz, 1H), 8.20 (s, 1H), 9.85 (s, 1H), 11.47 (s,1H).

Step 1g.N-(2-Chloro-6-methylphenyl)-2-(2-methyl-6-(piperazin-1-yl)pyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 109)

A mixture of compound 108 (2.60 g, 6.6 mmol), piperazine (5.60 g, 66.0mmol), potassium carbonate (1.82 g, 13.2 mmol) and DMF (15 mL) wasstirred at 135° C. for 12 hours. The solvent was evaporated under reducepressure and the residue was washed with water, acetone and ethylacetate in turn to obtain the title compound 109 as a pale yellow solid(1.8 g, 64%): LCMS: 444 [M+1]⁺.

Step 1h. Ethyl2-(4-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-yl)piperazin-1-yl)acetate(Compound 110-1)

A mixture of compound 109 (0.31 g, 0.70 mmol), ethyl 2-bromoacetate (117mg, 0.70 mmol), triethylamine (0.28 g, 0.70 mmol) and DMF (5 mL) wasstirred at 35° C. for 2 minutes. The solvent was evaporated under reducepressure to give the title compound 110-1 as a white solid (333 mg, 90%)which was used directly to the next step without further purification:LCMS: 530 [M+1]⁺.

Step 1i.N-(2-Chloro-6-methylphenyl)-2-(6-(4-(2-(hydroxyamino)-2-oxoethyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 1)

To a stirred solution of hydroxylamine hydrochloride (4.67 g, 67.0 mmol)in methanol (24 mL) at 0° C. was added a solution of potassium hydroxide(5.61 g, 100.0 mmol) in methanol (14 mL). After addition, the mixturewas stirred for 30 minutes at 0° C., and was allowed to stand at 0° C.The resulting precipitate was isolated, and the solution was prepared togive free hydroxylamine.

The above freshly prepared hydroxylamine solution (0.5 mL, 0.89 mmol)was placed in 5 mL flask. Compound 110-1 (333 mg, 0.63 mmol) was addedto this solution under ultrasonic for 10 minutes. The reaction processwas monitored by TLC. The mixture was neutralized with acetic acid andwas then concentrated under reduce pressure. The residue was purified bypreparative HPLC to give the title compound 1 as a white solid (50 mg,16%): LCMS: 517 [M+1]⁺; ¹H NMR (DMSO-d₆) δ 2.20 (s, 3H), 2.38 (s, 3H),2.58 (m, 4H), 2.90 (s, 2H), 3.51 (m, 4H), 6.02 (s, 1H), 7.26 (m, 2H),7.37 (m, 1H), 8.20 (s, 1H), 8.80 (s, 1H), 9.86 (s, 1H), 10.47 (s, 1H),11.46 (s, 1H).

Example 2 Preparation ofN-(2-chloro-6-methylphenyl)-5-(6-(4-(3-(hydroxyamino)-3-oxopropyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)-4H-pyrrole-3-carboxamide(Compound 2) Step 2a. Methyl3-(4-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-yl)piperazin-1-yl)propanoate(Compound 110-2)

The title compound 110-2 was prepared as a pale yellow solid (0.31 g,74%) from compound 109 (0.35 g, 0.79 mmol), methyl 3-bromopropanoate(0.13 g, 0.78 mmol), DIEA (0.21 g, 1.58 mmol) and DMF (5 mL) using aprocedure similar to that described for compound 110-1 (Example 1):LCMS: 530[M+1]⁺.

Step 2b.N-(2-Chloro-6-methylphenyl)-2-(6-(4-(3-(hydroxyamino)-3-oxopropyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 2)

The title compound 2 was prepared as a white solid (60 mg, 19%) fromcompound 110-2 (0.31 g, 0.59 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LCMS: 531 [M+1]⁺; ¹H NMR (DMSO-d₆)δ2.16 (t, J=6.9 Hz, 2H), 2.24 (s, 3H), 2.41 (s, 3H), 2.54 (m, 4H), 2.57(t, J=6.6 Hz, 2H), 3.50 (m, 4H), 6.05 (s, 1H), 7.25 (m, 2H), 7.37 (m,1H), 8.23 (s, 1H), 8.88 (s, 1H), 9.90 (s, 1H), 10.42 (s, 1H), 11.51 (s,1H).

Example 3 Preparation ofN-(2-chloro-6-methylphenyl)-2-(6-(4-(4-(hydroxyamino)-4-oxobutyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 3) Step 3a. Ethyl4-(4-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-yl)piperazin-1-yl)butanoate(Compound 110-3)

The title compound 110-3 was prepared as a pale yellow solid (0.22 g,71%) from compound 109 (0.25 g, 0.56 mmol), ethyl 4-bromobutanoate (0.12g, 0.56 mmol), DIEA (0.15 g, 0.56 mmol) and DMF (5 mL) using a proceduresimilar to that described for compound 110-1 (Example 1): LCMS: 558[M+1]⁺.

Step 3b.N-(2-chloro-6-methylphenyl)-2-(6-(4-(4-(hydroxyamino)-4-oxobutyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 3)

The title compound 3 was prepared as a white solid (30 mg, 14%) fromcompound 110-3 (0.22 g, 0.40 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LCMS: 545 [M+1]⁺; ¹H NMR (DMSO-d₆)δ1.69 (m, 2H), 2.01 (t, J=6.6 Hz, 2H), 2.25 (s, 3H), 2.30 (t, J=6.9,2H), 2.41 (m, 4H), 2.55 (s, 3H), 3.52 (m, 4H), 6.06 (s, 1H), 7.25 (m,2H), 7.36 (m, 1H), 8.23 (s, 1H), 8.70 (s, 1H), 9.90 (s, 1H), 10.37 (s,1H), 11.50 (s, 1H).

Example 4 Preparation ofN-(2-chloro-6-methylphenyl)-2-(6-(4-(5-(hydroxyamino)-5-oxopentyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 4) Step 4a. Methyl methyl5-(4-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-yl)piperazin-1-yl)pentanoate(Compound 110-4)

The title compound 110-4 was prepared as a pale yellow solid (120 mg,39%) from compound 109 (0.24 g, 0.54 mmol), methyl 5-bromopentanoate(0.12 g, 0.60 mmol), DIEA (1.54 g, 1.20 mmol) and DMF (3 mL) using aprocedure similar to that described for compound 110-1 (Example 1):LCMS: 558 [M+1]⁺.

Step 4b.N-(2-chloro-6-methylphenyl)-2-(6-(4-(5-(hydroxyamino)-5-oxopentyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 4)

The title compound 4 was prepared as a white solid (30 mg, 25%) fromcompound 110-4 (120 mg, 0.22 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LCMS: 559 [M+1]⁺; ¹H NMR (DMSO-d₆)δ1.44 (m, 4H), 1.95 (t, J=7.5 Hz, 2H), 2.22 (s, 3H), 2.26 (t, J=6.9 Hz,2H), 2.37 (m, 7H), 3.47 (m, 4H), 6.07 (s, 1H), 7.25 (m, 2H), 7.37 (dd,J=2.1 Hz, J=7.2 Hz, 2H), 8.23 (s, 1H), 9.93 (s, 1H).

Example 5 Preparation ofN-(2-chloro-6-methylphenyl)-2-(6-(4-(6-(hydroxyamino)-6-oxohexyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 5) Step 5a. Ethyl6-(4-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-yl)piperazin-1-yl)hexanoate(Compound 110-5)

The title compound 110-5 was prepared as a brown solid (120 mg, 41%)from compound 109 (0.22 g, 0.495 mol), ethyl 6-bromohexanoate (0.12 g,0.495 mmol), potassium carbonate (0.22 g, 1.60 mmol) and DMF (5 mL)using a procedure similar to that described for compound 110-1 (Example1): LCMS: 586 [M+1]⁺.

Step 5b.N-(2-Chloro-6-methylphenyl)-2-(6-(4-(6-(hydroxyamino)-6-oxohexyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 5)

The title compound 5 was prepared as a white solid (30 mg, 26%) fromcompound 110-5 (120 mg, 0.20 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LC-MS: 573 [M+1]⁺; ¹H NMR(DMSO-d₆) δ1.26 (m, 2H), 1.49 (m, 4H), 1.93 (t, J=7.2 Hz, 2H), 2.22 (s,3H), 2.26 (t, J=7.2 Hz, 2H), 2.48 (m, 7H), 3.47 (m, 4H), 6.04 (s, 1H),7.26 (m, 2H), 7.37 (m, 2H), 8.21 (s, 1H), 8.66 (s, 1H), 9.88 (s, 1H),10.33 (s, 1H), 10.33 (s, 1H).

Example 6 Preparation ofN-(2-chloro-6-methylphenyl)-2-(6-(2-(2-(hydroxyamino)-2-oxoethylamino)ethylamino)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide (Compound7) Step 6a.2-(6-(2-Aminoethylamino)-2-methylpyrimidin-4-ylamino)-N-(2-chloro-6-methylphenyl)thiazole-5-carboxamide(Compound 201)

A solution of compound 108 (3.0 g, 7.6 mmol) in ethane-1,2-diamine (50mL) was heated to 80° C. and stirred for 10 hours. The reaction was thenconcentrated under vacuum and the residue was partitioned between H₂Oand EtOAc. The EtOAc phase was separated, washed with brine, dried(Na₂SO₄) and concentrated under vacuum to yield the title compound 201as a brown solid (1.3 g, 40%). LC-MS: 418 [M+1]⁺, ¹H-NMR (DMSO-d₆): δ1.86 (s, 2H), 2.22 (s, 3H), 2.36 (s, 3H), 2.48 (t, J=6.0 Hz, 2H), 2.76(t, J=6.0 Hz, 2H), 3.15 (s, 1H), 5.88 (s, 1H), 7.26 (m, 2H), 7.37 (dd,J=2.4, J=6.9 Hz, 1H), 8.19 (s, 1H), 9.83 (s, 1H).

Step 6b. Ethyl2-(2-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-ylamino)ethylamino)acetate (Compound 202-7)

A solution of compound 201 (0.50 g, 1.2 mmol) in DMF (15 mL) was addedethyl 2-bromoacetate (0.2 g, 1.2 mmol) and K₂CO₃ (41 mg, 0.3 mmol). Thereaction was stirred at 30° C. for 2 hours. The mixture was concentratedunder vacuum and the residue was purified by column chromatograph toobtain title compound 202-7 as a pale yellow solid (110 mg, 22%): LC-MS:504 [M+1]⁺.

Step 6c.N-(2-Chloro-6-methylphenyl)-2-(6-(2-(2-(hydroxyamino)-2-oxoethylamino)ethylamino)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 7)

The title compound 7 was prepared as a pale yellow solid (42 mg, 37%)from compound 202-7 (110 mg, 0.29 mmol) using a procedure similar tothat described for compound 1 (Example 1): LC-MS: 491 [M+1]⁺, H-NMR(DMSO-d₆): δ 2.21 (s, 3H), 2.34 (s, 3H), 2.60 (t, J=6 Hz, 2H), 3.03 (s,2H), 3.11 (t, J=5.7 Hz, 2H), 5.86 (s, 1H), 7.22 (m, 2H), 7.36 (dd,J=2.1, J=7.2 Hz, 1H), 8.18 (s, 1H), 9.83 (s, 1H).

Example 7 Preparation ofN-(2-chloro-6-methylphenyl)-2-(6-(4-(7-(hydroxyamino)-7-oxoheptyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 6) Step 7a. ethyl7-(4-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-yl)piperazin-1-yl)heptanoate(Compound 110-6)

The title compound 110-6 was prepared as a brown solid (176 mg, 59%)from compound 109 (0.22 g, 0.50 mmol), ethyl 7-bromoheptanoate (0.12 g,0.506 mmol), diisopropylethylamine (0.13 g, 1.00 mmol) and DMF (5 mL)using a procedure similar to that described for compound 110-1 (Example1): LCMS: 600 [M+1]⁺.

Step 7b.N-(2-chloro-6-methylphenyl)-2-(6-(4-(6-(hydroxyamino)-7-oxoheptyl)piperazin-1-yl)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 6)

The title compound 6 was prepared as a white solid (32 mg, 82%) fromcompound 110-6 (40 mg, 0.067 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LC-MS: 587 [M+1]⁺; ¹H NMR(DMSO-d₆) δ1.24 (m, 4H), 1.44 (m, 4H), 1.92 (t, J=7.2 Hz, 2H), 2.22 (s,3H), 2.26 (t, J=6.3 Hz, 2H), 2.38 (ds, 7H), 3.48 (m, 4H), 6.03 (s, 1H),7.26 (m, 2H), 7.37 (m, 1H), 8.19 (s, 1H), 8.63 (ds, 1H), 9.83 (s, 1H),10.28 (s, 1H), 11.43 (s, 1H).

Example 8 Preparation ofN-(2-chloro-6-methylphenyl)-2-(6-(2-(3-(hydroxyamino)-3-oxopropylamino)ethylamino)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 8) Step 8a. Methyl3-(2-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-ylamino)ethylamino)propanoate(Compound 202-8)

The title compound 202-8 was prepared as a white solid (400 mg, 31%)from compound 201 (1.08 g, 2.6 mol), methyl 4-bromobutanoate (0.44 g,2.6 mmol) and K₂CO₃ (0.44 mg, 5.2 mmol) using a procedure similar tothat described for compound 202-7 (Example 6): LCMS 504 [M+1]⁺. ¹H-NMR((DMSO-d₆): δ 2.22 (s, 3H), 2.38 (s, 3H), 2.64 (t, J=6.9 Hz, 2H), 2.93(t, J=6.0 Hz, 2H), 3.03 (t, J=6.6 Hz, 2H), 3.61 (s, 3H), 7.26 (m, 3H),7.39 (m, 1H), 8.22 (s, 1H), 9.88 (s, 1H).

Step 8b.N-(2-chloro-6-methylphenyl)-2-(6-(2-(3-(hydroxyamino)-3-oxopropylamino)ethylamino)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 8)

The title compound 8 was prepared as a off white solid (30 mg, 60%) fromcompound 202-8 (51 mg, 0.10 mmol) using a procedure similar to thatdescribed for compound 1 (Example 1): LCMS 505 [M+1]⁺; ¹H NMR (DMSO-d₆),δ 2.13 (t, J=6.9 Hz

-   -   2H), 2.22 (s, 3H), 2.36 (s, 3H), 2.70 (t, J=6.6 Hz, 2H), 2.77        (t, J=6.9 Hz, 2H), 5.87 (s, 1H), 7.21 (m, 3H), 7.39 (m, 1H),        8.19 (s, 1H), 9.84 (s, 1H).

Example 9 Preparation ofN-(2-chloro-6-methylphenyl)-2-(6-(2-(6-(hydroxyamino)-6-oxohexylamino)ethylamino)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 11) Step 9a. Ethyl6-(2-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-ylamino)ethylamino)hexanoate(Compound 202-11)

The title compound 202-11 was prepared as a pale yellow solid (100 mg,17%) from compound 201 (0.50 g, 1.2 mol), ethyl ethyl 6-bromohexanoate(0.27 g, 1.2 mmol) and K₂CO₃ (41 mg, 0.3 mmol) using a procedure similarto that described for compound 202-7 (Example 6): H-NMR (CDCl₃): δ 1.24(m, 5H), 1.41 (m, 2H), 1.57 (m, 2H), 2.23 (t, J=7.2 Hz, 2H), 2.34 (s,3H), 2.50 (s, 3H), 2.57 (t, J=5.7 Hz, 2H), 2.84 (t, J=5.7 Hz, 2H), 3.37(m, 2H), 4.11 (q, J=7.2 Hz, 2H), 5.46 (ds, 1H), 5.70 (s, 1H), 7.16 (m,1H), 7.29 (m, 3H), 8.15 (s, 1H).

Step 9b.N-(2-chloro-6-methylphenyl)-2-(6-(2-(6-(hydroxyamino)-6-oxohexylamino)ethylamino)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 11)

The title compound 11 was prepared as a off white solid (34 mg, 33%)from compound 202-11 (100 mg, 0.18 mmol) using a procedure similar tothat described for compound 1 (Example 1): LCMS 547 [M+1]⁺; ¹H NMR(DMSO-d₆), δ 1.25 (m, 2H), 1.47 (m, 4H), 1.95 (t, J=7.2 Hz 2H), 2.21 (s,3H), 2.37 (s, 3H), 2.75 (t, J=6.9 Hz, 2H), 2.91 (t, J=6.6 Hz, 2H), 3.42(ds, 1H), 5.90 (s, 1H), 7.22 (m, 4H), 8.19 (s, 1H), 9.8 (s, 1H), 10.4(ds, 1H).

Example 10 Preparation ofN-(2-chloro-6-methylphenyl)-2-(6-(6-(hydroxyamino)-6-oxohexylamino)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 23) Step 10a.Methyl6-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-ylamino)hexanoate(Compound 301-23)

A solution of compound 108 (240 mg, 0.61 mmol), DMAC (15 mL), KOH (170mg, 3.05 mmol) and methyl 6-aminohexanoate (554 mg, 3.05 mmol) wasstirred for 12 h at 120° C. The reaction mixture was diluted with water,filtered and dried to give the crude compound 301-23 as a pale yellowpowder (88 mg, 30%) which was used directly to next step without furtherpurification. LCMS: 503 [M+1]⁺.

Step 10b.N-(2-chloro-6-methylphenyl)-2-(6-(6-(hydroxyamino)-6-oxohexylamino)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 23)

A mixture of compound 301-23 (88 mg, 0.18 mmol) and freshly preparedNH₂OH methanol solution (1.77 M, 2.10 mL) was stirred for 30 min at roomtemperature. The mixture was adjusted to pH=7.0 with AcOH and thesolvent was removed. The resulting residue was purified by columnchromatography to give the title compound 23 as a white powder (25 mg,29%): LCMS: 504 [M+1]⁺; ¹H NMR (DMSO-d₆) δ 11.30 (s, 1H), 10.29 (s, 1H),9.80 (s, 1H), 8.60 (s, 1H), 8.17 (s, 1H), 7.38 (dd, 1H, J=2.1, J=7.2Hz), 7.25 (m, 2H), 7.12 (m, 1H), 5.83 (s, 1H), 3.13 (brs, 2H), 2.34 (s,3H), 2.22 (s, 3H), 1.93 (m, 2H), 1.50 (m, 1H), 1.26 (m, 2H).

Example 11 Preparation ofN-(2-chloro-6-methylphenyl)-2-(6-(7-(hydroxyamino)-7-oxoheptylamino)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 24) Step 11a. Methyl7-(6-(5-(2-chloro-6-methylphenylcarbamoyl)thiazol-2-ylamino)-2-methylpyrimidin-4-ylamino)heptanoate(Compound 301-24)

The title compound 301-24 was prepared as a crude pale yellow solid (120mg, 38%) from compound 108 (240 mg, 0.61 mmol), DMAC (15 mL), KOH (170mg, 3.05 mmol) and methyl 7-aminoheptanoate (596 mg, 3.05 mmol) using aprocedure similar to that described for compound 301-23 (Example 10):LCMS: 517 [M+1]⁺.

Step 11b.N-(2-chloro-6-methylphenyl)-2-(6-(7-(hydroxyamino)-7-oxoheptylamino)-2-methylpyrimidin-4-ylamino)thiazole-5-carboxamide(Compound 24)

The title compound 24 was prepared as a white solid (35 mg, 30%) fromcompound 301-24 (120 mg, 0.23 mmol) and freshly prepared hydroxylaminemethanol solution (1.77 M, 3.28 mL) using a procedure similar to thatdescribed for compound 23 (Example 10): m.p. 150.7° C. (decomp.), LCMS:518 [M+1]⁺; ¹H NMR (DMSO-d₆) δ 11.37 (s, 1H), 10.33 (s, 1H), 9.85 (s,1H), 8.66 (s, 1H), 8.18 (s, 1H), 7.39 (dd, 1H, J=2.1, J=7.2 Hz), 7.26(m, 2H), 7.19 (m, 1H), 5.82 (s, 1H), 3.14 (brs, 2H), 2.34 (s, 3H), 2.22(s, 3H), 1.92 (m, 2H), 1.47 (m, 4H), 1.27 (m, 4H).

Biological Assays:

As stated hereinbefore the derivatives defined in the present inventionpossess anti-proliferation activity. These properties may be assessed,for example, using one of the procedures set out below:

(a) An In Vitro Assay which Determines the Ability of a Test Compound toInhibit a Tyrosine Kinase.

The ability of compounds to inhibit tyrosine kinase (Abl1, Src, c-Kit,and PDGFR-beta) activity is assayed using HTSCAN™ Receptor Kinase AssayKits (Cell Signaling Technologies, Danvers, Mass.). Abl1 tyrosine kinaseis obtained in partially purified form from GST-kinase fusion proteinwhich is produced using a baculovirus expression system from a constructexpressing human Abl1 (Pro118-Ser553) (GenBank Accession No.NM_(—)005157) with an amino-terminal GST tag. Src tyrosine kinase isobtained in partially purified form from GST-kinase fusion protein whichis produced using a baculovirus expression system from a constructexpressing full length human Src (Met1-Leu536) (GenBank Accession No.NM_(—)005417) with an amino-terminal GST tag. c-Kit tyrosine kinase isobtained in partially purified form from GST-kinase fusion protein whichis produced using a baculovirus expression system from a constructexpressing human c-Kit (Thr544-Val976) with an amino-terminal GST tag.PDGFR-beta tyrosine kinase was produced using a baculovirus expressionsystem from a construct containing a human PDGFR-beta c-DNA (GenBankAccession No. NM_(—)002609) fragment (Arg561-Leu1106) amino-terminallyfused to a GST-HIS6-Thrombin cleavage site. The proteins are purified byone-step affinity chromatography using glutathione-agarose. Ananti-phosphotyrosine monoclonal antibody, P-Tyr-100, is used to detectphosphorylation of biotinylated substrate peptides (Abl1 and Src,Biotin-Signal Transduction Protein (Tyr160); c-Kit, Biotinylated-KDR(Tyr996); PDGFR-β, Biotinylated-FLT3 (Tyr589)). Enzymatic activity istested in 60 mM HEPES, 5 mM MgCl2 5 mM MnCl2 200 μM ATP, 1.25 mM DTT, 3μM Na3VO4, 1.5 mM peptide, and 50 ng EGF Recpetor Kinase. Bound antibodyis detected using the DELFIA system (PerkinElmer, Wellesley, Mass.)consisting of DELFIA® Europium-labeled Anti-mouse IgG (PerkinElmer,#AD0124), DELFIA® Enhancement Solution (PerkinElmer, #1244-105), and aDELFIA® Streptavidin coated, 96-well Plate (PerkinElmer, AAAND-0005).Fluorescence is measured on a WALLAC Victor 2 plate reader and reportedas relative fluorescence units (RFU). Data are plotted using GraphPadPrism (v4.0a) and IC50's are calculated using a sigmoidal dose responsecurve fitting algorithm.

Test compounds are dissolved in dimethylsulphoxide (DMSO) to give a 20mM working stock concentration. Each assay is setup as follows: 100 μlof 10 mM ATP is added to 1.25 ml 6 mM substrate peptide. The mixture isdiluted with dH₂0 to 2.5 ml to make 2×ATP/substrate cocktail ([ATP]=400mM, [substrate]=3 mM). The enzyme is immediately transferred from −80°C. to ice. The enzyme is allowed to thaw on ice. The mixture ismicrocentrifuged briefly at 4° C. to bring liquid to the bottom of thevial and returned immediately to ice. 10 μl of DTT (1.25 mM) is added to2.5 ml of 4× HTScan™ Tyrosine Kinase Buffer (240 mM HEPES pH 7.5, 20 mMMgCl₂, 20 mM MnCl, 12 mM NaVO₃) to make DTT/Kinase buffer. 1.25 ml ofDTT/Kinase buffer is transferred to enzyme tube to make a 4× reactioncocktail ([enzyme]=4 ng/μL in 4× reaction cocktail). 12.5 μl of the 4×reaction cocktail is incubated with 12.5 μL/well of prediluted compoundof interest (usually around 10 μM) for 5 minutes at room temperature. 25μl of 2×ATP/substrate cocktail is added to 25 μl/well preincubatedreaction cocktail/compound. The reaction plate is incubated at roomtemperature for 30 minutes. 50 μl/well Stop Buffer (50 mM EDTA, pH 8) isadded to stop the reaction. 25 μl of each reaction and 75 μl dH₂O/wellis transferred to a 96-well streptavidin-coated plate and incubated atroom temperature for 60 minutes. The plate is washed three times with200 μl/well PBS/T (PBS, 0.05% Tween-20). The primary antibody,Phospho-Tyrosine mAb (P-Tyr-100), is diluted 1:1000 in PBS/T with 1%bovine serum albumin (BSA). 100 μl/well primary antibody is added andthe mixture is incubated at room temperature for 60 minutes. The platesare again washed three times with 200 μl/well PBS/T. Europium labeledanti-mouse IgG is diluted 1:500 in PBS/T with 1% BSA. 100 μl/welldiluted antibody is added and the mixture is incubated at roomtemperature for 30 minutes. The plate is washed five times with 200μl/well PBS/T. 100 μl/well DELFIA® Enhancement Solution is added and themixture is incubated at room temperature for 5 minutes. 615 nmfluorescence emission is detected using an appropriate Time-ResolvedPlate Reader.

(b) An In Vitro Assay which Determines the Ability of a Test Compound toInhibit HDAC Enzymatic Activity.

HDAC inhibitors is screened using an HDAC fluorimetric assay kit(AK-500, Biomol, Plymouth Meeting, Pa.). Test compounds are dissolved indimethylsulphoxide (DMSO) to give a 20 mM working stock concentration.Fluorescence is measured on a WALLAC Victor 2 plate reader and reportedas relative fluorescence units (RFU). Data are plotted using GraphPadPrism (v4.0a) and IC50's calculated using a sigmoidal dose responsecurve fitting algorithm.

Each assay is setup as follows: Defrost all kit components and kept onice until use. Dilute HeLa nuclear extract 1:29 in Assay Buffer (50 mMTris/Cl, pH 8.0, 137 mM NaCl, 2.7 mM KCl, 1 mM MgCl2). Prepare dilutionsof Trichostatin A (TSA, positive control) and tested compounds in assaybuffer (5× of final concentration). Dilute Fluor de Lys™ Substrate inassay buffer to 100 uM (50 fold=2× final). Dilute Fluor de Lys™developer concentrate 20-fold (e.g. 50 μl plus 950 μl Assay Buffer) incold assay buffer. Second, dilute the 0.2 mM Trichostatin A 100-fold inthe 1× Developer (e.g. 10 μl in 1 ml; final Trichostatin A concentrationin the 1× Developer=2 μM; final concentration after addition toHDAC/Substrate reaction=1 μM). Add Assay buffer, dilute trichostatin Aor test inhibitor to appropriate wells of the microtiter plate. Adddiluted HeLa extract or other HDAC sample to all wells except fornegative controls. Allow diluted Fluor de Lys™ Substrate and the samplesin the microtiter plate to equilibrate to assay temperature (e.g. 25 or37° C. Initiate HDAC reactions by adding diluted substrate (25 μl) toeach well and mixing thoroughly. Allow HDAC reactions to proceed for 1hour and then stopped them by addition of Fluor de Lys™ Developer (50μl). Incubate plate at room temperature (25° C.) for 10-15 min. Readsamples in a microtiter-plate reading fluorimeter capable of excitationat a wavelength in the range 350-380 nm and detection of emitted lightin the range 440-460 nm.

The following TABLE B lists compounds representative of the inventionand their activity in HDAC, SRC, c-Kit, PDGF and ABL assays. In theseassays, the following grading was used: I≧10 μM, 10 μM>II>1 μM, 1μM>III>0.1 μM, and IV≦0.1 μM for IC₅₀.

TABLE B Compound No. HDAC ABL SRC c-Kit PDGFb Lyn Lck 1 II IV IV IV 2 IIIV IV IV 3 II IV IV 4 III IV IV IV 5 IV IV IV IV IV IV 6 III IV IV IV IVIV 7 I IV IV 11 IV IV IV IV IV IV IV 23 IV IV IV IV IV IV IV 24 IV IV IVIV IV IV IV 30 I III III

The patent and scientific literature referred to herein establishes theknowledge that is available to those with skill in the art. All UnitedStates patents and published or unpublished United States patentapplications cited herein are incorporated by reference. All publishedforeign patents and patent applications cited herein are herebyincorporated by reference. All other published references, documents,manuscripts and scientific literature cited herein are herebyincorporated by reference.

While this invention has been particularly shown and described withreferences to preferred embodiments thereof, it will be understood bythose skilled in the art that various changes in form and details may bemade therein without departing from the scope of the inventionencompassed by the appended claims.

What is claimed is:
 1. A method of treating a cancer which issusceptible to inhibition of one or more of Src, Bcr-Abl and HDAC, in asubject in need thereof, the method comprising administering to thesubject a therapeutically effective amount of a compound represented byformula (I):

or a pharmaceutically acceptable salt or solvate thereof, wherein Cz ispyrimidine or substituted pyrimidine; Ar is phenyl or substitutedphenyl; X₃ is NH; Z₂ is S; Y₂ is N; R₂₁ is hydrogen; B is a direct bondor straight- or branched-, substituted or unsubstituted alkyl,substituted or unsubstituted alkenyl, substituted or unsubstitutedalkynyl, arylalkyl, arylalkenyl, arylalkynyl, heteroarylalkyl,heteroarylalkenyl, heteroarylalkynyl, heterocyclylalkyl,heterocyclylalkenyl, heterocyclylalkynyl, aryl, heteroaryl,heterocyclyl, cycloalkyl, cycloalkenyl, alkylarylalkyl,alkylarylalkenyl, alkylarylalkynyl, alkenylarylalkyl,alkenylarylalkenyl, alkenylarylalkynyl, alkynylarylalkyl,alkynylarylalkenyl, alkynylarylalkynyl, alkylheteroarylalkyl,alkylheteroarylalkenyl, alkylheteroarylalkynyl, alkenylheteroarylalkyl,alkenylheteroarylalkenyl, alkenylheteroarylalkynyl,alkynylheteroarylalkyl, alkynylheteroarylalkenyl,alkynylheteroarylalkynyl, alkylheterocyclylalkyl,alkylheterocyclylalkenyl, alkylheterocycloalkynyl,alkenylheterocyclylalkyl, alkenylheterocyclylalkenyl,alkenylheterocyclylalkynyl, alkynylheterocyclylalkyl,alkynylheterocyclylalkenyl, alkynylheterocyclylalkynyl, alkylaryl,alkenylaryl, alkynylaryl, alkylheteroaryl, alkenylheteroaryl, oralkynylhereroaryl, in which one or more methylenes can be interrupted orterminated by O, S, S(O), SO₂, N(R₈), C(O), substituted or unsubstitutedaryl, substituted or unsubstituted heteroaryl, substituted orunsubstituted heterocyclic; R₈ is hydrogen, acyl, aliphatic orsubstituted aliphatic; C is

where W is O; Y is absent; Z is N; R₇ is absent; and R₉ is OR′, whereinR′ is hydrogen.
 2. The method of claim 1, wherein the compound ofFormula I is represented by formula (III):

wherein R₄ is hydrogen, aliphatic or substituted aliphatic; and C, B,Y₂, Z₂, Ar and R₂₁ are as defined in claim
 1. 3. The method of claim 1,wherein the compound of Formula I is represented by formula (IV):

wherein is 1-9; and R′, Z₂, Ar and R₂₁ are as defined in claim
 1. 4. Themethod of claim 1, wherein the compound of Formula I is represented byformula (V):

wherein m and n are independently 1-9; R₁₈ is independently R₈; and R′,Z₂, Ar, R₈, and R₂₁ are as defined in claim
 1. 5. The method of claim 1,wherein the compound of Formula I is represented by formula (VI):

wherein m is 0-9; W is O, NH, alkylamino, S, SO, or SO₂; and Z₂, Ar, R′,R₈, and R₂₁ are as defined in claim
 1. 6. A method of treating a cancerwhich is susceptible to inhibition of one or more of Src, Bcr-Abl andHDAC, in a subject in need thereof, the method comprising administeringto the subject a therapeutically effective amount of a compoundrepresented by formula (VIII):

or a pharmaceutically acceptable salt or solvate thereof, wherein M₁ isabsent, 0, NH, alkylamino, C₁-C₆ alkyl, C₂-C₆ alkenyl, or C₂-C₆ alkynyl;M₂ is absent, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆ alkynyl, 0, NH,alkylamine, S, SO, SO₂, heterocyclic, heteroaryl, aryl or C═O; M₃ isabsent, O, NH, alkylamino, S, SO, SO₂, C₁-C₆ alkyl, C₂-C₆ alkenyl, C₂-C₆alkynyl, heterocyclic, heteroaryl, aryl or C═O; M₄ is absent, O, NH,alkylamino, CO, S, SO, SO₂, heterocyclic, heteroaryl or aryl; M₅ isabsent, C₁-C₈ alkyl, C₂-C₈ alkenyl, C₂-C₈ alkynyl, heterocyclic,heteroaryl or aryl; R′ is hydrogen; Z₂ is S; Ar is phenyl or substitutedphenyl; Y is absent; R₇ is absent; R₈ is hydrogen; and R₂₁ is hydrogen.7. The method of claim 1, wherein the cancer is leukemia, lymphoma or asolid tumor.
 8. The method of claim 1, wherein the cancer is selectedfrom the group consisting of cutaneous T-cell lymphoma, noncutaneousperipheral T-cell lymphoma, chronic myelogenous leukemia, acutemyelogenous leukemia, non-Hodgkin lymphoma, acute lymphatic leukemia,chronic lymphatic leukemia, hepatocellular carcinoma, head and neckcancers, renal cancer, ovarian cancer, lung cancer, breast cancer,pancreatic cancer, and stomach cancer.
 9. The method of claim 6, whereinthe cancer is leukemia, lymphoma or a solid tumor.
 10. The method ofclaim 6, wherein the cancer is selected from the group consisting ofcutaneous T-cell lymphoma, noncutaneous peripheral T-cell lymphoma,chronic myelogenous leukemia, acute myelogenous leukemia, non-Hodgkinlymphoma, acute lymphatic leukemia, chronic lymphatic leukemia,hepatocellular carcinoma, head and neck cancers, renal cancer, ovariancancer, lung cancer, breast cancer, pancreatic cancer, and stomachcancer.